Methods of treating subjects having clinically significant signs and symptoms associated with blood cell differentiation

Abstract

Disclosed herein are methods of treating a subject having clinically significant signs and symptoms associated with blood cell differentiation and treated with an agent that reduces the level and / or activity of BRG1 and / or BRM. The methods disclosed herein may comprise, for example, administering to the subject an effective amount of a corticosteroid, hydroxyurea, or furosemide, or performing leukocyte separation on the subject.

Description

Background Art

[0001] Chromatin regulation is critical for gene expression, and ATP-dependent chromatin remodeling is a mechanism by which this gene expression occurs. The human Switch / Sucrose Non-Fermentable (SWI / SNF) chromatin remodeling complex (also known as the BAF complex) has two SWI2-like ATPases, called BRG1 and BRM. The transcriptional activator BRG1, also known as the ATP-dependent chromatin remodeler SMARCA4, is encoded by the SMARCA4 gene on chromosome 19. BRG1 is overexpressed in some cancer tumors and is required for cancer cell proliferation. BRM, also known as the rational global transcriptional activator SNF2L2 and / or the ATP-dependent chromatin remodeler SMARCA2, is encoded by the SMARCA2 gene on chromosome 9 and has been shown to be critical for tumor cell growth in cells characterized by BRG1 loss-of-function mutations. Inactivation of BRG and / or BRM leads to downstream effects in cells, including cell cycle arrest and tumor suppression.

[0002] Chromatin regulation is essential for gene expression, and ATP-dependent chromatin remodeling is a mechanism by which this gene expression occurs. The human switch / sucrose non-fermentable (SWI / SNF) chromatin remodeling complex (also known as the BAF complex) has two SWI2-like ATPases, called BRG1 (Brahma-associated gene 1) and BRM (Brahma). The transcriptional activator BRG1, also known as the ATP-dependent chromatin remodeler SMARCA4, is encoded by the SMARCA4 gene on chromosome 19. BRG1 is overexpressed in some cancer tumors and is required for cancer cell proliferation. BRM, also known as the rational global transcriptional activator SNF2L2 and / or the ATP-dependent chromatin remodeler SMARCA2, is encoded by the SMARCA2 gene on chromosome 9 and has been shown to be essential for tumor cell growth in cells characterized by BRG1 loss-of-function mutations. Inactivation of BRG and / or BRM leads to downstream effects in cells, including cell cycle arrest and tumor suppression.

[0003] Hematologic cancers, also called blood cancers, are cancers that begin in blood-forming tissues (such as the bone marrow) or in cells of the immune system, such as leukemia. Leukemia is a cancer found in the blood and bone marrow that is caused by the rapid production of abnormal white blood cells. In most hematologic cancers, normal blood cell development is interrupted by the uncontrolled growth of abnormal types of blood cells. Abnormal blood cells prevent the blood from performing many of its functions. Hematologic cancers account for about 10% of all newly diagnosed cancers. The 5-year relative survival rate for hematologic cancers is about 50% to about 90%. Summary of the invention

[0004] The invention features methods of treating a subject treated with an agent that reduces the level and / or activity of BRG1 and / or BRM. The subject may have clinically significant signs and symptoms associated with blood cell differentiation, may have or be suspected of having a differentiation syndrome, or may have non-infectious leukocytosis. The methods described herein include administering to the subject an effective amount of a corticosteroid, hydroxyurea, or furosemide, or performing leukapheresis on the subject.

[0005] In one aspect, the invention provides methods of treating a subject having differentiation syndrome and being treated with an agent that decreases the level and / or activity of BRG1 and / or BRMs by administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

[0006] In another aspect, the invention provides methods of treating a subject suspected of having differentiation syndrome and being treated with an agent that decreases the level and / or activity of BRG1 and / or BRMs by administering to the subject an effective amount of corticosteroids, hydroxyurea or furosemide, or performing leukapheresis on the subject.

[0007] In some embodiments, the subject is treated with an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM of a leukemia (e.g., acute myeloid leukemia). In some embodiments, the subject is treated with an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM of a myelodysplastic syndrome (MDS).

[0008] In some embodiments, the method further comprises the step of administering an agent that decreases the level and / or activity of BRG1 and / or BRM.

[0009] In another aspect, the invention provides a method of treating a subject having leukemia or myelodysplastic syndrome by administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM and, if the subject has clinically significant signs and symptoms associated with blood cell differentiation, administering to the subject an effective amount of a corticosteroid, hydroxyurea, or furosemide, or performing leukapheresis on the subject.

[0010] In another aspect, the invention provides a method of treating a subject having leukemia or myelodysplastic syndrome by administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM and, if the subject has differentiation syndrome, administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

[0011] In another aspect, the present invention provides a method of treating a subject having leukemia or myelodysplastic syndrome by administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM, and if the subject is suspected of having differentiation syndrome, administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

[0012] In some embodiments, the subject has leukemia. In some embodiments, the leukemia is acute myeloid leukemia. In some embodiments, the leukemia is relapsed or refractory acute myeloid leukemia. In some embodiments, the subject has myelodysplastic syndrome.

[0013] In some embodiments, the subject exhibits one or more of the following symptoms: unexplained fever, rash, hypoxia, respiratory impairment, interstitial lung infiltrates, pleural and / or pericardial effusions, weight gain, renal failure, dyspnea, clinical deterioration, fluid in or around the lungs, fluid around the heart, leg swelling, increased bilirubin, and increased liver enzymes. In some embodiments, a blood sample from a subject includes an elevated absolute neutrophil count (ANC) and / or an elevated platelet count.

[0014] In some embodiments, an effective amount of corticosteroid is administered to the subject. In some embodiments, corticosteroid is systemically administered. In some embodiments, corticosteroid is administered orally or by injection. In some embodiments, a high dose regimen of corticosteroid is administered to the subject. In some embodiments, corticosteroid is administered to the subject for at least 3 days. In some embodiments, corticosteroid is dexamethasone, ethamethasone, hydrocortisone, cortisone, prednisone, prednisolone, methylprednisolone, triamcinolone, its pharmaceutically acceptable salt or a combination thereof. In some embodiments, if the clinically significant signs and symptoms associated with blood cell differentiation or the symptoms of differentiation syndrome continue for at least 48 hours after starting to administer corticosteroids, the administration of the agent that reduces the level and / or activity of BRG1 and / or BRM is interrupted. In some embodiments, administration of the agent that reduces the level and / or activity of BRG1 and / or BRM is discontinued if clinically significant signs and symptoms associated with blood cell differentiation or symptoms of differentiation syndrome persist for at least 3 days after initiation of corticosteroid administration.

[0015] In some embodiments, an effective amount of a diuretic is administered to the subject. In some embodiments, the diuretic is administered systemically. In some embodiments, the diuretic is administered orally or by injection. In some embodiments, a high-dose regimen of a diuretic is administered to the subject. In some embodiments, the diuretic is furosemide or a pharmaceutically acceptable salt thereof.

[0016] In some embodiments, an effective amount of furosemide is administered to a subject. In some embodiments, furosemide is administered systemically. In some embodiments, furosemide is administered orally or by injection. In some embodiments, furosemide is administered intramuscularly or intravenously.

[0017] In some embodiments, the subject has symptoms of non-infectious leukocytosis.

[0018] In some embodiments, the method comprises the step of administering to the subject an effective amount of hydroxyurea. In some embodiments, an effective amount of hydroxyurea is administered to the subject until the non-infectious leukocytosis improves or resolves.

[0019] In some embodiments, the method comprises the step of performing leukapheresis on the subject.

[0020] In some embodiments, the subject experiences hypervolemia. In some embodiments, an effective amount of furosemide is administered to the subject.

[0021] In some embodiments, the agent that decreases the level and / or activity of BRG1 and / or BRM is a compound of the following structure:

[0022]

[0023] or a pharmaceutically acceptable salt thereof.

[0024] In some embodiments, the agent that decreases the level and / or activity of BRG1 and / or BRM is a compound of the following structure:

[0025]

[0026] or a pharmaceutically acceptable salt thereof.

[0027] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is administered orally. In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is administered in a unit dosage form selected from the group consisting of a capsule or a tablet.

[0028] In some embodiments of any of the above aspects, the effective amount of the agent reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%) compared to a reference. In some embodiments, the effective amount of the agent reduces the level and / or activity of BRG1 by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%) compared to a reference. In some embodiments, the effective amount of the agent reduces the level and / or activity of BRG1 by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%).

[0029] In some embodiments, the effective amount of the agent reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In some embodiments, the effective amount of the agent reduces the level and / or activity of BRG1 by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or longer).

[0030] In some embodiments of any of the above aspects, the effective amount of the agent reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In some embodiments, the effective amount of the agent reduces the level and / or activity of a BRM by at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference. In some embodiments, the effective amount of the agent reduces the level and / or activity of a BRM by at least 90% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%).

[0031] In some embodiments, the effective amount of the agent reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 12 hours (e.g., 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 48 ​​hours, 72 hours, or more). In some embodiments, the effective amount of the agent reduces the level and / or activity of a BRM by at least 5% (e.g., 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) compared to a reference for at least 4 days (e.g., 5 days, 6 days, 7 days, 14 days, 28 days, or longer).

[0032] In some embodiments, the anti-cancer therapy and the agent that reduces the level and / or activity of BRG1 and / or BRM in cells are administered within 28 days of each other, and each is administered in an amount that together is effective to treat the subject.

[0033] In some embodiments, the subject or leukemia (e.g., acute myeloid leukemia) has and / or has been identified as having a BRG1 loss-of-function mutation. In some embodiments, the subject or leukemia (e.g., acute myeloid leukemia) has and / or has been identified as having a BRM loss-of-function mutation. In some embodiments, the leukemia (e.g., acute myeloid leukemia) comprises a BRG1 T910M mutation.

[0034] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is a small molecule compound, such as a small molecule BRG1 and / or BRM inhibitor. In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is a small molecule compound, such as a small molecule BRG1 inhibitor. In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is a small molecule compound, such as a small molecule BRM inhibitor or a degrader.

[0035] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula I or a pharmaceutically acceptable salt thereof:

[0036]

[0037] Where m is 0, 1, 2, 3 or 4;

[0038] X1 is N or CH; and

[0039] Each R 1 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino.

[0040] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula II or a pharmaceutically acceptable salt thereof:

[0041]

[0042] Where R 2 is phenyl substituted by hydroxy and optionally substituted by one or more groups independently selected from the group consisting of halo, cyano, trifluoromethyl, trifluoromethoxy, C 1-3 Alkyl and C 1-3 Alkoxy;

[0043] R 3 Selected from the group consisting of: -R a 、-OR a 、-N(R a )2, -S(O)2R a and -C(O)-N(R a )2;

[0044] Each R a are independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl, wherein each C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 The alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of: R b , oxo, halogen, -NO2, -N(R b )2、-CN、-C(O)-N(R b )2、-S(O)-N(R b )2、-S(O)2-N(R b )2.-OR b 、-SRb 、-OC(O)-R b 、-C(O)-R b 、-C(O)-OR b 、-S(O)-R b 、-S(O)2-R b 、-N(R b )-C(O)-R b 、-N(R b )-S(O)-R b 、-N(R b )-C(O)-N(R b )2 and -N(R b )-S(O)2-R b ;

[0045] Each R b are independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, 3-15 membered carbocyclic group and 3-15 membered heterocyclic group, wherein each C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted by one or more groups independently selected from Rc; or two Rb together with the nitrogen to which they are attached form a heterocyclyl, the heterocyclyl being optionally substituted by one or more groups independently selected from oxo, halo and C 1-3 alkyl, which is optionally substituted with one or more groups independently selected from the group consisting of oxo and halo;

[0046] Each Rc is independently selected from the group consisting of: oxo, halo, -NO2, -N(R d )2、-CN、-C(O)-N(R d )2、-S(O)-N(R d )2. -S(O) 2- N(R d )2. -SR d 、-OC(O)-R d 、-C(O)-R d 、-C(O)-OR d 、-S(O)-R d 、-S(O)2-R d 、-N(R d )-C(O)-R d 、-N(R d)-S(O)-R d 、-N(R d )-C(O)-N(R d )2、-N(R d )-S(O)2-R d , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 3-15 membered carbocyclic group and 3-15 membered heterocyclic group, any C1-6 alkyl, C 2-6 Alkenyl, C 2-6 The alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of: R d , oxo, halogen, -NO2, -N(R d )2、-CN、-C(O)-N(R d )2、-S(O)-N(R d )2、-S(O)2-N(R d )2.-OR d 、-SR d 、-OC(O)-R d 、-C(O)-R d 、-C(O)-R d 、-S(O)-R d 、-S(O)2-R d 、-N(R d )-C(O)-R d 、-N(R d )-S(O)-R d 、-N(R d )-C(O)-N(R d )2 and -N(R d )-S(O)2-R d ;

[0047] Each R d are independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, carbocyclic and carbocyclic (C 1-3 alkyl)-;

[0048] R 4 It is H, C 1-6 Alkyl or -C(=O)-C 1-6 Alkyl; and

[0049] R 5 Is H or C 1-6 alkyl.

[0050] The compound of formula II can be synthesized by methods known in the art, such as those described in U.S. Patent Publication No. 2018 / 0086720, the synthesis methods of which are incorporated by reference.

[0051] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula III or a pharmaceutically acceptable salt thereof:

[0052]

[0053] Where R 6 is halo, such as fluoro or chloro;

[0054] R 7 is hydrogen, optionally substituted amino or optionally substituted C 1-6 Alkyl; and

[0055] R 8 is an optionally substituted C 6-10 Aryl or optionally substituted C 2-9 Heteroaryl.

[0056] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of any one of Compounds 1-16, or a pharmaceutically acceptable salt thereof:

[0057]

[0058] In some embodiments, the small molecule compound or a pharmaceutically acceptable salt thereof is a degradation agent. In some embodiments, the degradation agent has a structure of Formula IV:

[0059] ALB

[0060] Formula IV

[0061] Wherein A is a BRG1 and / or BRM binding moiety; L is a linker; and B is a degradation moiety, or a pharmaceutically acceptable salt thereof. In some embodiments, the degradation moiety is a ubiquitin ligase moiety. In some embodiments, the ubiquitin ligase binding moiety comprises a Cereblon ligand, an IAP (inhibitor of apoptosis) ligand, a mouse double minute 2 homolog (MDM2), a hydrophobic tag, or a von Hippel-Lindau ligand or a derivative or analog thereof.

[0062] In some embodiments, A includes the structure of any of Formulas I-III or any of Compounds 1-16.

[0063] In some embodiments, the hydrophobic tag comprises diphenylmethane, adamantane, or tri-Boc arginine, i.e., the hydrophobic tag comprises the following structure:

[0064]

[0065] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula A:

[0066]

[0067] Where X 1 is CH2, O, S or NR 1 , where R 1 X is H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 2 is C=O, CH2 or R 3 and R 4 is independently H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; m is 0, 1, 2, 3, or 4; and each R 2 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino,

[0068] or a pharmaceutically acceptable salt thereof.

[0069] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0070]

[0071] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0072] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula B:

[0073]

[0074] Each R 4 , R 4’ and R 7 R is independently H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R 6 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; n is 0, 1, 2, 3 or 4; each R 8 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino; and each R 9 and R 10 are independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 Aryl, wherein R4' or R5 includes a bond to the linker, or a pharmaceutically acceptable salt thereof.

[0075] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0076]

[0077] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0078] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula C:

[0079]

[0080] Each R 11 , R 13 and R 15 R is independently H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 12 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R 14 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; p is 0, 1, 2, 3 or 4; each R 16 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino; q is 0, 1, 2, 3 or 4; and each R 17 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0081] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0082]

[0083] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0084] In some embodiments, the ubiquitin ligase binding moiety comprises a structure of Formula D:

[0085]

[0086] Each R 18 and R 19 are independently H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C10 Aryl; r1 is 0, 1, 2, 3 or 4; each R 20 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino; r2 is 0, 1, 2, 3 or 4; and each R 21 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0087] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0088]

[0089] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0090] In some embodiments, the linker has the structure of Formula V:

[0091] A 1- (B 1 ) f -(C 1 ) g -(B 2 ) h -(D)-(B 3 ) i -(C 2 ) j -(B 4 ) k -A 2

[0092] Formula V

[0093] Among them A 1 is the bond between the connector and A; A 2 is the bond between B and the joint; B 1 , B 2 , B 3 and B 4are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2 and NR N ; R N is hydrogen, optionally substituted C 1-4 Alkyl, optionally substituted C 2-4 Alkenyl, optionally substituted C 2-4 Alkynyl, optionally substituted C 2-6 Heterocyclic group, optionally substituted C 6-12 Aryl or optionally substituted C 1-7 Heteroalkyl; C 1 and C 2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl or phosphoryl; f, g, h, I, j and k are each independently 0 or 1; and D is an optionally substituted C 1-10 Alkyl, optionally substituted C 2-10 Alkenyl, optionally substituted C 2-10 Alkynyl, optionally substituted C 2-6 Heterocyclic group, optionally substituted C 6-12 Aryl, optionally substituted C2-C 10 Polyethylene glycol or optionally substituted C 1-10 Heteroalkyl, or connected to A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h - to - (B 3 ) i -(C 2 ) j -(B 4 ) k -A 2 chemical bonds.

[0094] In some embodiments, D is optionally substituted C2-C 10 In some embodiments, C 1 and C 2 Each is independently carbonyl or sulfonyl. 1 , B 2 , B 3 and B 4 are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2 and NR N RN is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 , B 2 , B3 and B 4 Each is independently selected from optionally substituted C1-C2 alkyl or optionally substituted C1-C3 heteroalkyl. In some embodiments, j is 0. In some embodiments, k is 0. In some embodiments, j and k are each independently 0. In some embodiments, f, g, h and i are each independently 1.

[0095] In some embodiments, the linker of Formula V has the structure of Formula Va:

[0096]

[0097] Among them A 1 is the bond between the linker and A, and A 2 It is the bond between B and the linker.

[0098] In some embodiments, D is optionally substituted C 1-10 In some embodiments, C 1 and C 2 In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2 and NR N , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 , B 2 , B 3 and B 4 Each independently selected from optionally substituted C1-C2 alkyl, O, S, S(O)2 and NR N , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 and B 4 Each is independently an optionally substituted C1-C2 alkyl. 1 and B 4 Each is independently C1 alkyl. 2 and B 4 Each is independently NR N , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 2 and B 4In some embodiments, f, g, h, I, j, and k are each independently 1.

[0099] In some embodiments, the linker of Formula V has the structure of Formula Vb:

[0100]

[0101] Among them A 1 is the bond between the linker and A, and A 2 It is the bond between B and the linker.

[0102] In some embodiments, the leukemia (e.g., acute myeloid leukemia) is resistant to one or more chemotherapeutic or cytotoxic agents (e.g., the leukemia (e.g., acute myeloid leukemia) has been determined to be resistant to a chemotherapeutic or cytotoxic agent (e.g., by a genetic marker), or is likely to be resistant to a chemotherapeutic or cytotoxic agent (e.g., a leukemia (e.g., acute myeloid leukemia) that is unresponsive to a chemotherapeutic or cytotoxic agent). In some embodiments, the leukemia (e.g., acute myeloid leukemia) is unresponsive to one or more chemotherapeutic or cytotoxic agents. In some embodiments, the leukemia (e.g., acute myeloid leukemia) is resistant to dacarbazine, temozolomide, cisplatin, treosulfan, fotemustine, IMCgp100, CTLA-4 inhibitors (e.g., ipilimumab), PD-1 inhibitors (e.g., Nivolumab or pembrolizumab), PD- The patient is resistant or unresponsive to L1 inhibitors (e.g., atezolizumab, avelumab, or durvalumab), mitogen-activated protein kinase (MEK) inhibitors (e.g., selumetinib, binimetinib, or tametinib), and / or protein kinase C (PKC) inhibitors (e.g., sotrastaurin or LXS196, also known as IDE196).

[0103] In some embodiments, the leukemia (e.g., acute myeloid leukemia) is resistant or unresponsive to a previously administered therapeutic agent (e.g., a MEK inhibitor or a PKC inhibitor) for treating uveal melanoma. For example, in some embodiments, the leukemia (e.g., acute myeloid leukemia) is resistant or unresponsive to a mitogen-activated protein kinase (MEK) inhibitor (e.g., selumetinib, bimetinib, or tumetinib) and / or a protein kinase C (PKC) inhibitor (e.g., sotrastuzumab or LXS196).

[0104] In some embodiments, the agent that decreases the level and / or activity of BRG1 and / or BRM in a cell is a small molecule compound, an antibody, an enzyme, and / or a polynucleotide.

[0105] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is an enzyme, such as a clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (e.g., CRISPR-associated protein 9 (Cas9), CRISPR-associated protein 12a (Cas12a), zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), or a meganuclease).

[0106] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is a polynucleotide, such as an antisense nucleic acid, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), a micro RNA (miRNA), a CRISPR / Cas 9 nucleotide, or a ribozyme.

[0107] In some embodiments, the leukemia is acute myeloid leukemia.

[0108] In some embodiments, the leukemia is advanced.

[0109] Chemical terms

[0110] For any of the following chemical definitions, the number following the atomic symbol represents the total number of atoms of the element present in the particular chemical moiety. It should be understood that other atoms, such as hydrogen atoms or substituents as described herein, may be present as needed to satisfy the valence of the atoms. For example, an unsubstituted C2 alkyl group has the formula -CH2CH3. When used with the groups defined herein, references to the number of carbon atoms include the divalent carbons in acetal and ketal groups, but do not include the carbonyl carbon in acyl, ester, carbonate or carbamate groups. References to the number of oxygen, nitrogen or sulfur atoms in a heteroaryl group include only those atoms that form part of a heterocyclic ring.

[0111] As used herein, the term "acyl" means a hydrogen or alkyl group connected to a parent molecular group by a carbonyl group as defined herein, and is exemplified by formyl (i.e., carboxyaldehyde), acetyl, trifluoroacetyl, propionyl, and butyryl. Exemplary unsubstituted acyl groups include 1 to 6 carbons, 1 to 11 carbons, or 1 to 21 carbons.

[0112] As used herein, the term "alkyl" refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon group having 1 to 20 carbon atoms (eg, 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms).

[0113] Alkylene is a divalent alkyl group. The term "alkenyl" as used herein alone or in combination with other groups refers to a straight or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 or 2 carbon atoms).

[0114] The term "alkynyl" as used herein alone or in combination with other groups refers to a straight or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6 or 2 carbon atoms).

[0115] As used herein, the term "amino" refers to -N(R N1 )2, where each R N1 are independently H, OH, NO2, N(R N2 2. SO2OR N2 、SO2R N2 , SOR N2 , N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or other acyl described herein), wherein these listed R N1 Each of the groups may be optionally substituted; or both R N1 are combined to form an alkylene group or a heteroalkylene group, and wherein each R N2 The amino group of the compounds described herein can be an unsubstituted amino group (ie, -NH2) or a substituted amino group (ie, -N(R N1 )2).

[0116] As used herein, the term "aryl" refers to an aromatic mono- or poly-carbocyclic group of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl and 1H-indenyl.

[0117] As used herein, the term "arylalkyl" refers to an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups are 7 to 30 carbon atoms (e.g., 7 to 16 carbon atoms or 7 to 20 carbon atoms, e.g., C1-C6 alkyl, C6-C 10 Aryl, C1-C 10 Alkyl C6-C 10 Aryl or C1-C 20 Alkyl C6-C 10 In some embodiments, the alkyl and aryl groups may each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups.

[0118] As used herein, the term "azido" refers to a -N3 group.

[0119] As used herein, the term "bridged polycycloalkyl" refers to a bridged polycyclic group of 5 to 20 carbons containing 1 to 3 bridges.

[0120] As used herein, the term "cyano" refers to a -CN group.

[0121] As used herein, the term "carbocyclyl" refers to a non-aromatic C3-C4 ring formed by carbon atoms. 12 Monocyclic, bicyclic or tricyclic structures. Carbocyclic structures include cycloalkyl and unsaturated carbocyclic groups.

[0122] As used herein, the term "cycloalkyl" refers to a saturated, non-aromatic, monovalent, mono- or poly-carbocyclic group of 3 to 10, preferably 3 to 6 carbon atoms. This term is further exemplified by groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl and adamantyl.

[0123] As used herein, the term "halogen" means a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) group.

[0124] As used herein, the term "heteroalkyl" refers to an alkyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkyl group may be further substituted by 1, 2, 3 or 4 substituents as described herein for the alkyl group. An example of a heteroalkyl group is an "alkoxy group", which, as used herein, refers to an alkyl-O- (e.g., methoxy and ethoxy). Heteroalkylene is a divalent heteroalkyl group. As used herein, the term "heteroalkenyl" refers to an alkenyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkenyl group may be further substituted by 1, 2, 3 or 4 substituents as described herein for the alkenyl group. An example of a heteroalkenyl group is an "alkenyloxy group", which, as used herein, refers to an alkenyl-O-. Heteroalkenylene is a divalent heteroalkenyl group. As used herein, the term "heteroalkynyl" refers to an alkynyl group as defined herein in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen or sulfur. In some embodiments, the heteroalkynyl group can be further substituted with 1, 2, 3, or 4 substituents as described herein for alkynyl groups. An example of a heteroalkynyl group is "alkynyloxy," which, as used herein, refers to alkynyl-O-. A heteroalkynylene group is a divalent heteroalkynyl group.

[0125] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or polycyclic group of 5 to 12 atoms having at least one aromatic ring and containing 1, 2 or 3 ring atoms selected from nitrogen, oxygen and sulfur, the remaining ring atoms being carbon. One or two ring carbon atoms of the heteroaryl may be replaced by a carbonyl group. Examples of heteroaryl are pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxazolyl and thiazolyl.

[0126] As used herein, the term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups are 7 to 30 carbon atoms (e.g., 7 to 16 carbon atoms or 7 to 20 carbon atoms, such as C1-C6 alkyl, C2-C9 heteroaryl, C1-C 10 Alkyl C2-C9 heteroaryl or C1-C 20 In some embodiments, alkyl and heteroaryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups.

[0127] As used herein, the term "heterocyclyl" refers to a mono- or polycyclic group having from 3 to 12 atoms having at least one ring containing 1, 2, 3 or 4 ring atoms selected from N, O or S, wherein no ring is aromatic. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl and 1,3-dioxanyl.

[0128] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups are 7 to 30 carbon atoms (e.g., 7 to 16 carbon atoms or 7 to 20 carbon atoms, such as C1-C6 alkyl, C2-C9 heterocyclyl, C1-C 10 Alkyl C2-C9 heterocyclic or C1-C 20 In some embodiments, alkyl and heterocyclyl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the corresponding groups.

[0129] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with an -OH group.

[0130] As used herein, the term "hydroxy" refers to an -OH group.

[0131] As used herein, the term "N-protecting group" means those groups intended to protect amino groups from undesired reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis," 3rd Edition (John Wiley & Sons, New York, 1999). N-protecting groups include, but are not limited to, acyl, aroyl or carbamoyl groups, such as formyl, acetyl, propionyl, pivaloyl, tert-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl and chiral auxiliaries, such as protected or unprotected D, L or D, L-amino acids, such as alanine, leucine and phenylalanine; sulfonyl-containing groups, such as benzenesulfonyl and p-toluenesulfonyl; carbamate-forming groups, such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, methoxybenzyloxycarbonyl, 2,4-20-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylcarbonyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, diphenylmethyloxycarbonyl, tert-butoxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl and phenylthiocarbonyl; arylalkyl groups such as benzyl, triphenylmethyl and benzyloxymethyl; and silyl groups such as trimethylsilyl. Preferred N-protecting groups are allyloxycarbonyl, formyl, acetyl, benzoyl, pivaloyl, tert-butylacetyl, alanyl, phenylsulfonyl, benzyl, tert-butoxycarbonyl (Boc) and benzyloxycarbonyl (Cbz).

[0132] As used herein, the term "nitro" refers to a -NO2 group.

[0133] As used herein, the term "thiol" refers to a -SH group.

[0134] Alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclic radical (e.g., cycloalkyl), aryl, heteroaryl and heterocyclic radical can be substituted or unsubstituted. Unless otherwise indicated, when substituted, there are generally 1 to 4 substituents. Substituents include, for example, alkyl (e.g., unsubstituted and substituted, wherein the substituent includes any group described herein, such as aryl, halo, hydroxyl), aryl (e.g., substituted and unsubstituted phenyl), carbocyclic radical (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluorine), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy or thioalkoxy), heteroaryl, heterocyclic radical, amino (e.g., NH or mono- or dialkylamino), azido, cyano, nitro or thiol. Aryl, carbocyclic (eg, cycloalkyl), heteroaryl, and heterocyclic groups may also be substituted with alkyl (unsubstituted and substituted alkyl, such as arylalkyl (eg, substituted and unsubstituted benzyl)).

[0135] Compounds described herein may have one or more asymmetric carbon atoms, and may exist as optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates, or mixtures of diastereomeric racemates. Optically active forms may be obtained, for example, by resolving racemates, by asymmetric synthesis or asymmetric chromatography (chromatography using chiral adsorbents or eluents). That is, certain disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in spatial arrangement. Enantiomers are paired stereoisomers whose mirror images are non-superimposable, and the most common reason is that they contain asymmetrically substituted carbon atoms that act as chiral centers. "Enantiomers" means one of a pair of molecules that are mirror images of each other and non-superimposable. Diastereomers are stereoisomers that are independent of mirror images, and the most common reason is that they contain two or more asymmetrically substituted carbon atoms, and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared, for example, by separating enantiomers from racemates using one or more well-known techniques and methods, such as chiral chromatography and separation methods based on chiral chromatography. One skilled in the art can readily determine the appropriate techniques and / or methods for separating enantiomers of the compounds described herein from a racemic mixture. "Racemate" or "racemic mixture" refers to a compound containing two enantiomers, wherein such a mixture does not exhibit optical activity; that is, they do not rotate the plane of polarized light. "Geometric isomers" refers to isomers that differ in the orientation of substituent atoms associated with a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. The atoms on each side of the carbon-carbon double bond (except H) may be in E (substituents on 25 opposite sides of the carbon-carbon double bond) or Z (substituents on the same side) configuration. "R", "S", "S* ”、“R *"," E," "Z," "cis," and "trans" represent configurations relative to the core molecule. Certain disclosed compounds may exist as atropisomers. Atropisomers are stereoisomers resulting from hindered rotation about a single bond, where the steric strain barrier to rotation is sufficiently high to allow separation of conformers. The compounds described herein can be prepared as individual isomers by isomer-specific synthesis or resolved from an isomer mixture. Conventional resolution techniques include forming salts of the free bases of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming salts of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), using optical The isomeric mixtures of the compounds disclosed herein are prepared by forming esters or amides of each isomer of an isomeric pair with a chemically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or by resolving isomeric mixtures of the starting materials or final products using various well-known chromatographic methods. When the stereochemistry of a disclosed compound is named or depicted by structure, the stereoisomer named or depicted is at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the enantiomer depicted or named is at least 60%, 70%, 80%, 90%, 99%, or 99.9% optically pure by weight. . When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. The percent optical purity is the ratio of the weight of the enantiomer relative to the weight of the enantiomer plus the weight of its optical isomers. The diastereomeric purity by weight is the ratio of the weight of one diastereomer to the weight of all diastereomers. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by mole fraction. The percent purity by mole fraction is the ratio of the number of moles of enantiomer to the number of moles of enantiomer plus the number of moles of its optical isomers. Similarly, the percent purity by mole fraction is the ratio of the number of moles of diastereomer to the number of moles of diastereomer plus the number of moles of its isomers.When a disclosed compound is named or depicted by structure without indicating stereochemistry and the compound has at least one chiral center, it is understood that the name or structure encompasses either enantiomer of the compound without the corresponding optical isomer, a racemic mixture of the compound, or a mixture enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating stereochemistry and has two or more chiral centers, it is understood that the name or structure encompasses a diastereomer without other diastereomers, a plurality of diastereomers without other diastereomer pairs, a mixture of diastereomers, a mixture of diastereomer pairs, a mixture of diastereomers in which one diastereomer is enriched relative to one or more other diastereomers, or a mixture of diastereomers in which one or more diastereomers are enriched relative to other diastereomers. The present invention encompasses all of these forms.

[0136] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Methods and materials for use in the present disclosure are described herein; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, the present specification, including definitions, shall prevail.

[0137] definition

[0138] In this application, unless the context clearly indicates otherwise, (i) the term "a / a" may be understood to mean "at least one / a"; (ii) the term "or" may be understood to mean "and / or"; and (iii) the terms "comprising" and "including" may be understood to cover the listed components or steps, whether they are presented by themselves or together with one or more other components or steps.

[0139] As used herein, the terms "about" and "approximately" refer to values ​​within 10% above or below the stated value. For example, the term "about 5%" means a range from 4.5% to 5.5%.

[0140] As used herein, the term "administering" refers to administering a composition (e.g., a compound or a formulation comprising a compound as described herein) to a subject or system. It can be administered to an animal subject (e.g., to a human) by any appropriate route. For example, in some embodiments, administration can be bronchial (including by bronchial instillation), buccal, enteral, intradermal, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intratumoral, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal, and vitreous.

[0141] As used herein, the term "BAF complex" refers to BRG 1 or HRBM-associated factor complex in human cells.

[0142] As used herein, the term "BAF complex-associated disorder" refers to a disorder caused or affected by the activity level of the BAF complex.

[0143] As used herein, the term "BRG1" refers to the ATP-dependent chromatin remodeling factor SMARCA4. BRG1 is a component of the BAF complex, which is a SWI / SNF ATPase chromatin remodeling complex. Human BRG1 is encoded by the SMARCA4 gene on chromosome 19, the nucleic acid sequence of which is set forth in SEQ ID NO: 1 (GenBank Accession No.: NM_001128849.1 (mRNA); www.ncbi.nlm.nih.gov / nuccore / NM_001128849.1?report=fasta).

[0144]

[0145]

[0146]

[0147] The term "BRG1" also refers to natural variants of wild-type human BRG1 protein, such as proteins that have at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% or more identity) to the amino acid sequence of wild-type BRG1, the amino acid sequence of which is set forth in SEQ ID NO: 2 (UniProt Accession No.: P51532; www.uniprot.org / uniprot / P51532.fasta).

[0148] SEQ ID NO: 2.

[0149]

[0150] As used herein, the term "BRM" refers to the rational global transcriptional activator SNF2L2. BRM is a component of the BAF complex, which is a SWI / SNF ATPase chromatin remodeling complex. Human BRM is encoded by the SMARCA2 gene on chromosome 9, the nucleic acid sequence of which is set forth in SEQ ID NO: 3 (GenBank Accession No.: NM_003070.4; www.ncbi.nlm.nih.gov / nuccore / NM_003070.4?report=fasta).

[0151] SEQ ID NO: 3.

[0152]

[0153]

[0154] The term "BRM" also refers to natural variants of wild-type human BRM protein, such as proteins that have at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% identity or more) to the amino acid sequence of wild-type BRM, the amino acid sequence of which is set forth in SEQ ID NO:4 (Uniprot Accession No.: P51531; www.uniprot.org / uniprot / P51531.fasta).

[0155] SEQ ID NO: 4.

[0156]

[0157] As used herein, the term "BRG1 activity" refers to the BRG1 enzyme ATPase activity.

[0158] As used herein, the term "BRG1 loss-of-function mutation" refers to a mutation in BRG1 that results in reduced protein activity (e.g., a reduction in BRG1 activity of at least 1%, such as a reduction in BRG1 activity of 2%, 5%, 10%, 25%, 50%, or 100%). Exemplary BRG1 loss-of-function mutations include, but are not limited to, homozygous BRG1 mutations and deletions of the C-terminus of BRG1.

[0159] As used herein, the term "BRG1 loss-of-function disorder" refers to a disorder (e.g., a leukemia (e.g., acute myeloid leukemia)) manifested by a decrease in BRG1 activity (e.g., a decrease in BRG1 activity of at least 1%, e.g., a decrease in BRG1 activity of 2%, 5%, 10%, 25%, 50%, or 100%).

[0160] As used herein, "combination therapy" or "combined administration" means administering two (or more) different agents or treatments to a subject as part of a treatment regimen defined for a specific disease or illness. The treatment regimen defines the dosage and administration cycle of each agent so that the effects of a single agent on the subject overlap. In some embodiments, the delivery of the two or more agents is simultaneous or concurrent, and the agent can be co-formulated. In some embodiments, the two or more agents are not co-formulated and are administered in a sequential manner as part of a prescription regimen. In some embodiments, the administration of two or more agents or treatments in a combined form causes a reduction in symptoms or other parameters associated with the disorder greater than the reduction observed in a single agent or treatment delivered or in the absence of the agent. The effects of the two treatments may be partially cumulative, completely cumulative, or greater than cumulative (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent may be achieved by any appropriate route including, but not limited to, oral routes, intravenous routes, intramuscular routes, and direct absorption by mucosal tissues. The therapeutic agent may be administered by the same route or by different routes. For example, the first therapeutic agent in the combination may be administered by intravenous injection, while the second therapeutic agent in the combination may be administered orally.

[0161] As used herein, the term "CTLA-4 inhibitor" refers to a compound (such as an antibody) that is capable of inhibiting the activity of a protein encoded by the CTLA4 gene in humans. Known CTLA-4 inhibitors include ipilimumab.

[0162] By "reduced level" or "increased level" of a protein or RNA is meant that the level of the protein or RNA is decreased or increased, respectively, compared to a reference (e.g., decreased or increased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more; decreased or increased by about 10%, about 150%, about 200%, about 300%, about 400%, about 500% or more, compared to a reference). The level of a protein may be expressed as a percentage of mass / vol (e.g., g / dL, mg / mL, μg / nL, ng / mL) or relative to the total protein in the sample.

[0163] By "reducing the activity of the BAF complex" is meant reducing the level of activity associated with the BAF complex, or a related downstream effect. A non-limiting example of reducing the activity of the BAF complex is Sox2 activation. The activity level of the BAF complex can be measured using any method known in the art, such as the method described in Kadoch et al., Cell, 2013, 153, 71-85, the methods of which are incorporated herein by reference.

[0164] As used herein, the term "degrader" refers to a small molecule compound that includes a degradation moiety, wherein the compound interacts with a protein (e.g., BRG1 and / or BRM) in a manner that results in degradation of the protein, e.g., binding of the compound results in a decrease in the level of the protein by at least 5%, e.g., in a cell or subject.

[0165] As used herein, the term "degradation moiety" refers to a moiety whose binding causes degradation of a protein (eg, BRG1 and / or BRM). In one embodiment, the moiety binds to a protease or ubiquitin ligase that metabolizes a protein (eg, BRG1 and / or BRM).

[0166] As used herein, a leukemia (e.g., acute myeloid leukemia) "determined to be resistant" refers to a leukemia (e.g., acute myeloid leukemia) that is resistant based on lack of or reduced responsiveness to chemotherapeutic agents, or a leukemia predicted to be resistant based on a prognostic assay (e.g., a gene expression assay).

[0167] By "drug resistant" is meant a leukemia (eg, acute myeloid leukemia) that is unresponsive or exhibits a reduced response to one or more chemotherapeutic agents (eg, any of the agents described herein).

[0168] As used herein, the term "not responsive to a prior therapy" or "refractory to a prior therapy" refers to a leukemia (eg, acute myeloid leukemia) that has progressed despite treatment with the therapy.

[0169] "Reducing the activity of BRG1 and / or BRM" means reducing the activity level associated with BRG1 and / or BRM, or the associated downstream effects. A non-limiting example of inhibiting the activity of BRG1 and / or BRM is reducing the level of BAF complex (e.g., GBAF) in a cell. The activity level of BRG1 and / or BRM can be measured using any method known in the art. In some embodiments, the agent that reduces the activity of BRG1 and / or BRM is a small molecule BRG1 and / or BRM inhibitor.

[0170] "Decreasing the level of BRG1 and / or BRM" means reducing the level of BRG1 and / or BRM in a cell or a subject. The level of BRG1 and / or BRM can be measured using any method known in the art.

[0171] As used herein, the term "inhibit BRG and / or BRM" refers to blocking or reducing the level or activity of the ATPase catalytic binding domain or bromodomain of a protein. BRG1 and / or BRM inhibition can be determined using methods known in the art, such as BRG and / or BRM ATPase assays, Nano DSF assays, or BRG1 and / or BRM luciferase cell assays.

[0172] "Level" means the level of a protein or mRNA encoding the protein compared to a reference. A reference can be any useful reference as defined herein. A "reduced level" or "increased level" of a protein means that the protein level is reduced or increased (e.g., reduced or increased by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more compared to a reference; a reduction or increase of more than about 1% compared to a reference. The level of protein can be expressed as mass / vol (e.g., g / dL, mg / mL, μg / mL, ng / mL) or as a percentage relative to the total protein or mRNA in the sample.

[0173] As used herein, the term "inhibitor" refers to any agent that reduces the level and / or activity of a protein (eg, BRG1 and / or BRM). Non-limiting examples of inhibitors include small molecule inhibitors, degraders, antibodies, enzymes, or polynucleotides (eg, siRNA).

[0174] As used herein, the term "LXS196," also known as IDE196, refers to a PKC inhibitor having the following structure:

[0175]

[0176] or a pharmaceutically acceptable salt thereof.

[0177] As used herein, the term "MEK inhibitor" refers to a compound that is capable of inhibiting the activity of mitogen-activated protein kinase MEK1 or MEK2. The MEK inhibitor may be, for example, selumetinib, bimetinib or tumetinib.

[0178] As used herein, the terms "effective amount," "therapeutically effective amount," and "sufficient amount" of an agent described herein that reduces the level and / or activity of BRG1 and / or BRM (e.g., in a cell or subject) refers to an amount sufficient to produce a beneficial or desired result (including a clinical result) when administered to a subject, including a human, and thus "effective amount" or its synonyms depend on the context in which it is used. For example, in the context of treating v, the amount of an agent that reduces the level and / or activity of BRG1 and / or BRM is sufficient to achieve a therapeutic response compared to the response obtained without the administration of the agent that reduces the level and / or activity of BRG1 and / or BRM. The amount of a given agent that will correspond to such an amount of reducing the level and / or activity of BRG1 and / or BRM described herein will vary depending on various factors, such as a given agent, pharmaceutical formulation, route of administration, type of disease or condition, identity of the subject being treated (e.g., age, sex, and / or weight) or identity of the host, etc., but can still be routinely determined by those skilled in the art. Likewise, as used herein, a "therapeutically effective amount" of an agent that reduces the level and / or activity of BRG1 and / or BRMs disclosed herein is an amount that results in a beneficial or desired outcome in a subject compared to a control. As defined herein, a therapeutically effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRMs disclosed herein can be readily determined by one of ordinary skill in the art by conventional methods known in the art. The dosage regimen can be adjusted to provide the optimal therapeutic response.

[0179] The term "inhibitory RNA agent" refers to an RNA or its analog that has sufficient sequence complementarity to the target RNA to guide RNA interference. Examples also include DNA that can be used to prepare RNA. RNA interference (RNAi) refers to a sequence-specific or selective process by which a target molecule (e.g., a target gene, protein, or RNA) is downregulated. In general, interfering RNA ("iRNA") is a double-stranded short interfering RNA (siRNA), a short hairpin RNA (shRNA), or a single-stranded microRNA (miRNA) that causes catalytic degradation of a specific mRNA, and can also be used to reduce or inhibit gene expression.

[0180] The terms "short interfering RNA" and "siRNA" (also referred to as "small interfering RNA") refer to RNA agents of about 10-50 nucleotides in length, preferably double-stranded agents, the strands optionally having overhangs, the overhangs comprising, for example, 1, 2 or 3 overhanging nucleotides (or nucleotide analogs), which are capable of directing or mediating RNA interference. Naturally occurring siRNAs are produced by the RNAi machinery of cells (e.g., Dicer or its homologs) from longer dsRNA molecules (e.g., length>25 nucleotides).

[0181] As used herein, the term "shRNA" refers to an RNA agent having a stem-loop structure, comprising a first and a second region of complementary sequence, the degree and orientation of complementarity of the regions being sufficient to allow base pairing to occur between the regions, the first and second regions being connected by a loop region, the loop being generated by the lack of base pairing between nucleotides (or nucleotide analogs) within the loop region.

[0182] The terms "miRNA" and "microRNA" refer to RNA agents of about 10-50 nucleotides, preferably between about 15-25 nucleotides in length, preferably single-stranded agents, which can guide or mediate RNA interference. Naturally occurring miRNAs are produced by Dicer from stem-loop precursor RNAs (i.e., pre-miRNAs). As used herein, the term "Dicer" includes Dicer and any Dicer orthologs or homologs that can process dsRNA structures into siRNA, miRNA, siRNA-like molecules, or miRNA-like molecules. Based on the fact that it has been found that naturally occurring miRNAs are expressed in a temporal manner (e.g., during development), the term microRNA ("miRNA") can be used interchangeably with the term "small temporal RNA" ("stRNA").

[0183] As used herein, the term "antisense" refers to a nucleic acid comprising a polynucleotide that is sufficiently complementary to all or a portion of a gene, primary transcript, or processed mRNA to interfere with the expression of an endogenous gene (e.g., BRG1 and / or BRM). "Complementary" polynucleotides are those that are capable of base pairing according to the standard Watson-Crick complementarity rules. Specifically, purines will base pair with pyrimidines, forming a combination of guanine and cytosine pairing (G:C) and adenine and thymine pairing (A:T) in the case of DNA, or adenine and uracil pairing (A:U) in the case of RNA. It should be understood that even if two polynucleotides are not completely complementary to each other, they can hybridize to each other as long as each polynucleotide has at least one region that is substantially complementary to each other.

[0184] The term "antisense nucleic acid" includes single-stranded RNA expression cassettes and double-stranded DNA expression cassettes that can be transcribed to produce antisense RNA. "Active" antisense nucleic acid is an antisense RNA molecule that can selectively hybridize with the primary transcript or mRNA of the coded polypeptide, and the polypeptide has at least 80% sequence identity (e.g., 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% or higher identity) with the targeted polypeptide sequence (e.g., BRG1 and / or BRM polypeptide sequence). Antisense nucleic acids can be complementary to the entire coding chain, or only to a portion thereof. In some embodiments, antisense nucleic acid molecules are antisense to the "coding region" of the coding chain of the nucleotide sequence. The term "coding region" refers to the region of the nucleotide sequence that contains codons that are translated into amino acid residues. In some embodiments, antisense nucleic acid molecules are antisense to the "non-coding region" of the coding chain of the nucleotide sequence. The term "non-coding region" refers to the 5' and 3' sequences that are not translated into amino acids and are located flanking the coding region (i.e., also referred to as 5' and 3' untranslated regions). Antisense nucleic acid molecules can be complementary to the entire coding region of an mRNA, or can be antisense to only a portion of the coding or non-coding region of an mRNA. For example, antisense oligonucleotides can be complementary to the region around the translation start site. The length of the antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides.

[0185] " Percentage of sequence identity (%) " with respect to reference polynucleotide or peptide sequence is defined as after comparing sequence and introducing room to realize maximum sequence identity percentage when necessary, the percentage of nucleic acid or amino acid identical with the nucleic acid in reference polynucleotide or peptide sequence in candidate sequence.Comparison for determining nucleic acid or amino acid sequence identity percentage can be realized in various ways within the capabilities of those skilled in the art, for example, using publicly available computer software, such as BLAST, BLAST-2 or Megalign software.Those skilled in the art can determine the parameters applicable to the comparison sequence, including any algorithm required for the maximum comparison in the total length of the compared sequence.For example, sequence comparison computer program BLAST can be used to generate sequence identity percentage value.For example, given nucleic acid or amino acid sequence A to, with or for given nucleic acid or amino acid sequence B sequence identity percentage (can also be alternatively described as given nucleic acid or amino acid sequence A to, with or for given nucleic acid or amino acid sequence B certain sequence identity percentage) is calculated as follows:

[0186] 100×(fraction X / Y)

[0187] wherein X is the number of nucleotides or amino acids scored as identical matches by a sequence alignment program (e.g., BLAST) in the program's alignment of A and B, and wherein Y is the total number of nucleic acids in B. It will be understood that when the length of nucleic acid or amino acid sequence A is not equal to the length of nucleic acid or amino acid sequence B, the percent sequence identity of A to B will not be equal to the percent sequence identity of B to A.

[0188] As used herein, the term "pharmaceutical composition" refers to a composition containing a compound as described herein, which is formulated with a pharmaceutically acceptable excipient and is suitable for administration to a mammal, such as a human. Typically, a pharmaceutical composition is manufactured or sold under the approval of a government regulatory agency as part of a therapeutic regimen for treating a mammalian disease. The pharmaceutical composition may be formulated, for example, for oral administration in a unit dosage form (e.g., tablets, capsules, caplets, soft capsules, or syrups); for topical administration (e.g., in the form of a cream, gel, lotion, or ointment); for intravenous administration (e.g., in the form of a sterile solution without microparticle plugs and in a solvent system suitable for intravenous use); or any other pharmaceutically acceptable formulation.

[0189] As used herein, "pharmaceutically acceptable excipients" refers to any ingredient other than the compounds described herein and having substantially non-toxic and non-inflammatory properties in patients (e.g., a vehicle capable of suspending or dissolving the active compound). Excipients may include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), softeners, emulsifiers, fillers (diluents), film formers or coatings, flavoring agents, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water of hydration.

[0190] As used herein, the term "pharmaceutically acceptable salt" means any pharmaceutically acceptable salt of any compound described herein. For example, pharmaceutically acceptable salts of any compound described herein include those salts that are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction, and commensurate with a reasonable benefit / risk ratio within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66: 1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (P. H. Stahl and C. G. Wermuth, eds.), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or prepared separately by reacting the free base with a suitable organic acid.

[0191] The compounds described herein may have ionizable groups so that they can be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids, or in the case of acidic forms of the compounds described herein, the salts may be prepared from inorganic or organic bases. Typically, the compounds are prepared or used as pharmaceutically acceptable salts, which are prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparing appropriate salts are well known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0192] As used herein, the term "PKC inhibitor" refers to a compound that is capable of inhibiting the activity of protein kinase C. The PKC inhibitor may be, for example, sotrastaurin or IDE196.

[0193] "Proliferation" as used in this application relates to the duplication or propagation of similar forms (of cells) arising from the constituent (cell) elements.

[0194] "Reference" means any useful reference for comparing protein or RNA levels. A reference can be any sample, standard, standard curve or level for comparison purposes. A reference can be a normal reference sample or reference standard or level. A "reference sample" can be, for example, a control, such as a predetermined negative control value, such as a "normal control" or a previous sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (e.g., a cell or tissue) from a subject without a disease; a sample from a subject diagnosed with a disease but not yet treated with the compounds of the invention; a sample from a subject treated with the compounds of the invention; or a sample of a purified protein or RNA (e.g., any of those described herein) at a known normal concentration. "Reference standard or level" means a value or value derived from a reference sample. A "normal control value" is a predetermined value indicating a non-disease state, for example, a value expected in a healthy control subject. Typically, a normal control value is expressed as a range ("between X and Y"), a high threshold ("not higher than X"), or a low threshold ("not lower than X"). A subject whose measured value is within the normal control value for a particular biomarker is generally referred to as being "within normal limits for that biomarker". A normal reference standard or level can be a value or value derived from a normal subject not suffering from a disease or condition, such as a leukemia (e.g., acute myeloid leukemia); a value or value derived from a subject who has been treated with a compound of the invention. In a preferred embodiment, the reference sample, standard or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, stage of disease, and general health. A standard curve of purified protein or RNA levels (e.g., any purified protein or RNA described herein) within the normal reference range can also be used as a reference.

[0195] As used herein, the term "subject" refers to any organism to which a composition according to the invention may be administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may be seeking or in need of treatment, requiring treatment, being treated, about to be treated, or a person or animal under the care of a trained professional for a particular disease or illness.

[0196] As used herein, the terms "treat," "treated," or "treating" refer to therapeutic treatment or any measure whose purpose is to alleviate (lessen) an unwanted physiological condition, disorder, or disease or to obtain a beneficial or desired clinical result. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; reduction in the extent of the disorder, condition, or disease; stabilization of the state of the disorder, condition, or disease (i.e., not worsening); delay in the onset or slowing of the progression of the disorder, condition, or disease; improvement or remission (whether partial or total) of the disorder, condition, or disease state; improvement in at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of the disorder, condition, or disease. Treatment includes inducing a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival compared to the expected survival if not receiving treatment. The compounds of the invention may also be used to "prophylactically" treat or "prevent" a disorder, for example, in a subject at increased risk of developing the disorder.

[0197] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Methods and materials for use in the present disclosure are described herein; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, the present specification, including definitions, shall prevail.

[0198] The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and claims. DETAILED DESCRIPTION

[0199] In general, the present invention provides methods for treating subjects having clinically significant signs and symptoms associated with blood cell differentiation and treated with an agent that reduces the level and / or activity of BRG1 and / or BRM. It has been unexpectedly discovered that some subjects may develop clinically significant signs and symptoms associated with blood cell differentiation (e.g., differentiation syndrome) when receiving an agent (e.g., for the treatment of leukemia (e.g., acute myeloid leukemia)) of the following structure:

[0200]

[0201] Without wishing to be bound by theory, it is believed that reduction in the level and / or activity of BRG1 and / or BRM (e.g., using the compounds described above) implies an interaction between the SPI1 transcription factor and BAF, thereby affecting myeloid cell differentiation, and in turn leading to the appearance of clinically significant signs and symptoms associated with blood cell differentiation. Thus, subjects treated with an agent that reduces the level and / or activity of BRG1 and / or BRM and having clinically significant signs and symptoms associated with blood cell differentiation can be treated using the methods described herein.

[0202] method

[0203] Methods disclosed herein can be used to treat subjects with clinically significant signs and symptoms associated with blood cell differentiation. For example, a subject may be suspected of having differentiation syndrome. Clinically significant signs and symptoms (such as symptoms of differentiation syndrome) may be one or more of the following: unexplained fever, rash, hypoxia, respiratory damage, interstitial lung infiltration, pleural and / or pericardial effusion, weight gain, renal failure, dyspnea, clinical deterioration, effusion in or around the lungs, effusion around the heart, leg swelling, bilirubin increase and liver enzyme increase. Typically, changes in blood samples of subjects treated with agents that reduce the level and / or activity of BRG1 and / or BRM can be monitored, such as changes in absolute neutrophil count (ANC), platelet count or white blood cell (WBC) count. Therefore, alternatively or in addition, blood samples from subjects can be tested to determine absolute neutrophil count (ANC) and / or platelet count. An increase in ANC of at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%) caused by therapy with an agent that reduces the level and / or activity of BRG1 and / or BRM may be a clinically significant sign or symptom associated with blood cell differentiation (e.g., a symptom that may be a differentiation syndrome). An increase in platelet count of at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%) caused by therapy with an agent that reduces the level and / or activity of BRG1 and / or BRM may be a clinically significant sign or symptom associated with blood cell differentiation (e.g., a symptom that may be a differentiation syndrome). The subject may also or alternatively have non-infectious leukocytosis (e.g., if the white blood cell (WBC) count is greater than 25x10 9 / L (for example, greater than 30x10 9 / L), or if the absolute increase in total WBC from baseline is greater than 15x10 9Any clinically significant signs and symptoms associated with blood cell differentiation (e.g., symptoms of differentiation syndrome) may be treatment-induced (e.g., treatment with an agent that reduces the level and / or activity of BRG1 and / or BRM).

[0204] Generally, therapy for clinically significant signs and symptoms associated with blood cell differentiation (eg, symptoms of differentiation syndrome) can be initiated at the onset of any of the clinically significant signs and symptoms associated with blood cell differentiation (eg, symptoms of differentiation syndrome).

[0205] The method of the present invention can include administration (preferably systemic administration) of corticosteroids (e.g., glucocorticoids; e.g., high-dose corticosteroids, e.g., high-dose glucocorticoids). The high-dose regimen of corticosteroids is well known in the art. For example, a high-dose systemic corticosteroid regimen can be intravenous administration of dexamethasone, e.g., with a dosage of 10 mg, e.g., every 12 hours. For example, a high-dose systemic corticosteroid regimen can be intravenous or intramuscular administration of prednisone, prednisolone, methylprednisolone, or a pharmaceutically acceptable salt thereof, e.g., with a dosage of at least 40 mg (e.g., 40 mg to 60 mg or 40 mg to 125 mg), e.g., daily. Typically, methylprednisolone is administered as a pharmaceutically acceptable salt of methylprednisolone succinate. Corticosteroid therapy (e.g., systemic corticosteroids, e.g., high dose, systemic corticosteroids) can continue for at least 3 days (e.g., until signs and symptoms subside).

[0206] The methods of the present invention may include administering (e.g., orally administering) hydroxyurea (e.g., to a subject with symptoms of non-infectious leukocytosis). Hydroxyurea may be administered, for example, at a dose of at least 2 grams (e.g., 2 to 4 grams, or 2 to 3 grams), for example, twice a day or three times a day.

[0207] The methods of the present invention may include administering (preferably systemically) a diuretic (e.g., furosemide). For example, the systemic regimen of the diuretic may be the intravenous or intramuscular administration of furosemide, for example at a dose of 20 mg or 40 mg, for example, slowly administered over 1-2 minutes. For example, the systemic regimen of the diuretic may be the intravenous or intramuscular administration of furosemide or a pharmaceutically acceptable salt thereof, for example at a dose of at least 20 mg (e.g., 20 mg to 40 mg). For example, the second dose of the diuretic may be the intravenous or intramuscular administration of furosemide or a pharmaceutically acceptable salt thereof, for example at a dose of at least 20 mg (e.g., 20 mg to 80 mg). For example, the systemic regimen of the diuretic may be the oral administration of furosemide, for example, at a dose of at least 20 mg (e.g., 20 mg to 80 mg).

[0208] If clinically significant signs and symptoms associated with blood cell differentiation or symptoms of differentiation syndrome persist for at least 48 hours (e.g., at least 3 days) after the start of corticosteroid administration, the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM may be discontinued. If clinically significant signs and symptoms associated with blood cell differentiation or symptoms of differentiation syndrome persist for at least 48 hours (e.g., at least 3 days) after the start of hydroxyurea administration, the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM may be discontinued. If clinically significant signs and symptoms associated with blood cell differentiation or symptoms of differentiation syndrome persist for at least 48 hours (e.g., at least 3 days) after the start of furosemide administration, the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM may be discontinued. If clinically significant signs and symptoms associated with blood cell differentiation or symptoms of differentiation syndrome persist for at least 48 hours (e.g., at least 3 days) after leukapheresis is performed on the subject, the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM may be discontinued.

[0209] If administration of an agent that reduces the level and / or activity of BRG1 and / or BRM is interrupted, its administration may be resumed if, for example, clinically significant signs and symptoms associated with blood cell differentiation (e.g., symptoms of differentiation syndrome) improve, e.g., to Grade 2 or less. Grade 1 differentiation syndrome is mild differentiation syndrome, Grade 2 differentiation syndrome is moderate differentiation syndrome, Grade 3 differentiation syndrome is severe differentiation syndrome, and Grade 4 differentiation syndrome is life-threatening differentiation syndrome.

[0210] While not being limited by theory, it is believed that compounds useful as agents for reducing the level and / or activity of BRG1 and / or BRM exert their ability to modulate the level, state and / or activity of the BAF complex by inhibiting the activity of BRG1 and / or BRM proteins within the BAF complex in a mammal. BAF complex-associated disorders include, but are not limited to, disorders associated with BRG1 loss-of-function mutations.

[0211] Compared to an untreated subject population, treating leukemia (e.g., acute myeloid leukemia) according to the methods described herein can result in an increase in the average survival time of a subject population treated according to the present invention. For example, the average survival time increases by more than 30 days (more than 60 days, 90 days, or 120 days). The increase in the average survival time of a population can be measured in any reproducible manner. The increase in the average survival time of a population can be measured, for example, by calculating the average survival length of a population after starting treatment with a compound of the present invention. The increase in the average survival time of a population can also be measured, for example, by calculating the average survival length of a population after completing a first round of treatment using a pharmaceutically acceptable salt of the present invention.

[0212] Compared with the untreated group, the treatment of leukemia (e.g., acute myeloid leukemia) can also result in a reduction in the mortality rate of the treated subject group. For example, the mortality rate is reduced by more than 2% (e.g., more than 5%, 10% or 25%). The reduction in the group mortality rate of the treated subject can be measured by any repeatable method, for example, by calculating the average number of disease-related deaths per unit time after the group begins to use the pharmaceutically acceptable salt of the present invention for treatment. The reduction in the group mortality rate can also be measured, for example, by calculating the average number of disease-related deaths per unit time after the group completes the first round of treatment using the pharmaceutically acceptable salt of the present invention.

[0213] BRG1 and / or BRM reducing agents

[0214] The agent described herein that reduces the level and / or activity of BRG1 and / or BRM in a cell can be an antibody, a protein (e.g., an enzyme), a polynucleotide, or a small molecule compound. The agent reduces the activity level or related downstream effects associated with BRG1 and / or BRM in a cell or subject, or reduces the level of BRG1 and / or BRM.

[0215] In some embodiments, the agent that decreases the level and / or activity of BRG1 and / or BRM in a cell is an enzyme, a polynucleotide, or a small molecule compound, such as a small molecule BRG1 and / or BRM inhibitor.

[0216] Antibody

[0217] The agent that reduces the level and / or activity of BRG1 and / or BRM can be an antibody or an antigen-binding fragment thereof. For example, the agent that reduces the level and / or activity of BRG1 and / or BRM described herein is an antibody that reduces or blocks the activity and / or function of BRG1 and / or BRM by binding to BRG1 and / or BRM.

[0218] The preparation and use of therapeutic antibodies against target antigens (e.g., BRG1 and / or BRM) are known in the art. For methods of preparing recombinant antibodies, including antibody engineering, the use of degenerate oligonucleotides, 5′-RACE, phage display and mutagenesis; antibody testing and characterization; antibody pharmacokinetics and pharmacodynamics; antibody purification and storage; and screening and labeling techniques, see, for example, the references cited above, and Zhiqiang An (ed.), Therapeutic Monoclonal Antibodies: From Bench to Clinic. 1st edition, Wiley 2009, and Greenfield (ed.), Antibodies: A Laboratory Manual. (2nd edition) Cold Spring Harbor Laboratory Press 2013.

[0219] Polynucleotide

[0220] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is a polynucleotide. In some embodiments, the polynucleotide is an inhibitory RNA molecule that works, for example, by means of RNA interference (RNAi) approach. Inhibitory RNA molecules can reduce the expression level (e.g., protein level or mRNA level) of BRG1 and / or BRM. For example, inhibitory RNA molecules include short interfering RNA (siRNA), short hairpin RNA (shRNA) and / or microRNA (miRNA) targeting full-length BRG1 and / or BRM. siRNA is a double-stranded RNA molecule that is generally about 19-25 base pairs in length. shRNA is an RNA molecule that includes a hairpin bend that reduces target gene expression by RNAi. MicroRNA is a non-coding RNA molecule that is generally about 22 nucleotides in length. MiRNA binds to the target site on the mRNA molecule and silences mRNA, for example, by causing the cutting of mRNA, the destabilization of mRNA or the inhibition of mRNA translation. Degradation is caused by enzymes, RNA-induced silencing complexes (RISC).

[0221] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is an antisense nucleic acid. Antisense nucleic acids include antisense RNA (asRNA) and antisense DNA (asDNA) molecules that are generally about 10 to 30 nucleotides in length, which recognize polynucleotide target sequences or sequence portions by hydrogen bond interactions with the nucleotide bases of the target sequence (e.g., BRG1 and / or BRM). The target sequence can be a single-stranded or double-stranded RNA, or a single-stranded or double-stranded DNA.

[0222] In some embodiments, the polynucleotide reduces the level and / or activity of a negative function modulator or a positive function modulator. In other embodiments, the polynucleotide reduces the level and / or activity of an inhibitor of a positive function modulator.

[0223] The polynucleotides of the present invention can be modified, for example, to contain modified nucleotides, such as 2'-fluoro, 2'-o-methyl, 2'v deoxy, non-locked nucleic acids, 2'-hydroxyl, thiophosphate, 2'-thiouridine, 4'-thiouridine, 2'-deoxyuridine. Without being limited to theory, it is believed that certain modifications can increase nuclease resistance and / or serum stability, or reduce immunogenicity. The polynucleotides mentioned above can also be provided in a specific form, such as liposomes, microspheres, or can be applied to gene therapy, or can be provided in combination with a connecting portion. Such connecting portions include polycations, such as polylysine, which acts as a charge neutralizer for a phosphate backbone, or hydrophobic portions that enhance interaction with a cell membrane or increase nucleic acid uptake, such as lipids (e.g., phospholipids, cholesterol, etc.). These portions can be connected to the 3' end or 5' end of a nucleic acid, and can also be connected by bases, sugars, or intramolecular nucleosidic bonds. Other portions can be capping groups specifically located at the 3' end or 5' end of a nucleic acid to prevent degradation by nucleases (e.g., exonucleases, RNases, etc.). Such end-capping groups include hydroxyl protecting groups known in the art, including glycols (e.g., polyethylene glycol and tetraethylene glycol). The inhibitory effect of polynucleotides can be examined in vivo and in vitro using the gene expression system based on cell lines or animals of the present invention. In some embodiments, the polynucleotides reduce the level and / or activity or function of BRG1 and / or BRM. In embodiments, the polynucleotides inhibit the expression of BRG1 and / or BRM. In other embodiments, the polynucleotides increase the degradation of BRG1 and / or BRM and / or reduce the stability (i.e., half-life) of BRG1 and / or BRM. The polynucleotides can be chemically synthesized or transcribed in vitro.

[0224] Inhibitory polynucleotides can be designed by methods well known in the art. Programs known in the art (including but not limited to those maintained on the websites of Thermo Fisher Scientific, German Cancer Research Center, and The Ohio State University Wexner Medical Center) can be used to design siRNA, miRNA, shRNA, and asRNA molecules with sufficient homology to provide unique sequence specificity required for degradation of any RNA. Systematic testing of several design species for optimizing inhibitory polynucleotide sequences can be routinely performed by those skilled in the art. Considerations when designing interfering polynucleotides include but are not limited to biophysical, thermodynamic, and structural considerations, base preferences at specific positions in the sense strand, and homology. The preparation and use of inhibitory therapeutic agents based on non-coding RNA (e.g., ribozymes, RNAse P, siRNA, and miRNA) are also known in the art, such as described in Sioud, RNA Therapeutics: Function, Design, and Delivery (Methods in Molecular Biology). Humana Press 2010. Table 1 below provides exemplary inhibitory polynucleotides for use in the methods of the present invention. In some embodiments, the inhibitory polynucleotide has a nucleic acid sequence that has at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to the nucleic acid sequence of an inhibitory polynucleotide in Table 1. In some embodiments, the inhibitory polynucleotide has a nucleic acid sequence that has at least 70% sequence identity (e.g., 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9% or more identity) to the nucleic acid sequence of an inhibitory polynucleotide in Table 1.

[0225] The construction of the vector for expressing the polynucleotide used in the present invention can be completed using conventional techniques, which do not require detailed explanation to those of ordinary skill in the art. In order to produce an effective expression vector, it is necessary to have a regulatory sequence for controlling the expression of the polynucleotide. These regulatory sequences include promoter and enhancer sequences, and are affected by specific cytokines that interact with these sequences, and are well known in the art.

[0226] Gene Editing

[0227] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is a component of a gene editing system. For example, the agent that reduces the level and / or activity of BRG1 and / or BRM introduces changes (e.g., insertions, deletions (e.g., knockouts), translocations, inversions, single point mutations, or other mutations) in BRG1 and / or BRM. In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is a nuclease. Exemplary gene editing systems include zinc finger nucleases (ZFNs), nucleases (TALENs) based on transcription activator-like effectors, and clustered regularly spaced short palindromic repeats (CRISPR) systems. Methods based on ZFNs, TALENs, and CRISPRs are described in, for example, Gaj et al., Trends Biotechnol. 31(7): 397-405(2013).

[0228] CRISPR refers to a group of clustered regularly interspaced short palindromic repeats (or a system comprising a group of clustered regularly interspaced short palindromic repeats). A CRISPR system refers to a system derived from CRISPR and Cas (CRISPR-associated protein) or other nucleases, which can be used to silence or mutate the genes described herein. The CRISPR system is a naturally occurring system found in bacterial and archaeal genomes. The CRISPR locus consists of alternating repeat sequences and spacer sequences. In a naturally occurring CRISPR system, the spacer is typically a sequence exogenous to the bacteria (e.g., a plasmid or phage sequence). The CRISPR system has been modified for gene editing in eukaryotes (e.g., altering, silencing, and / or enhancing certain genes). See, e.g., Wiedenheft et al., Nature 482(7385): 331-338 (2012). For example, such a modification of the system includes introducing a plasmid containing a specially designed CRISPR and one or more appropriate Cas proteins into a eukaryotic cell. The CRISPR locus is transcribed into RNA and processed by the Cas protein into small RNAs comprising a repeat sequence flanked by a spacer. Depending on the spacer sequence, the RNA acts as a guide to direct the Cas protein to silence a specific DNA / RNA sequence. See, e.g., Horvath et al., Science 327(5962):167-170(2010); Makarova et al., Biology Direct 1:7(2006); Pennisi, Science 341(6148):833-836(2013). In some embodiments, the CRISPR system includes a Cas9 protein, which is a nuclease that cuts both strands of DNA. See, e.g., supra.

[0229] In some embodiments, in the CRISPR system used as described herein, e.g., according to one or more methods described herein, the spacer of CRISPR is derived from a target gene sequence, e.g., derived from a BRG1 and / or BRM sequence. In some embodiments, in the CRISPR system used as described herein, e.g., according to one or more methods described herein, the spacer of CRISPR is derived from a target gene sequence, e.g., derived from a BRG1 sequence. In some embodiments, in the CRISPR system used as described herein, e.g., according to one or more methods described herein, the spacer of CRISPR is derived from a target gene sequence, e.g., derived from a BRM sequence.

[0230] In some embodiments, agents that reduce the level and / or activity of BRG1 and / or BRM include guide RNA (gRNA) in a CRISPR system for gene editing. Table 1 below provides exemplary gRNAs for the methods of the present invention. In embodiments, agents that reduce the level and / or activity of BRG1 and / or BRM include ZFNs or mRNAs encoding ZFNs that target (e.g., cut) nucleic acid sequences (e.g., DNA sequences) of BRG1 and / or BRM. In embodiments, agents that reduce the level and / or activity of BRG1 and / or BRM include TALENs or mRNAs encoding TALENs that target (e.g., cut) nucleic acid sequences (e.g., DNA sequences) of BRG1 and / or BRM. In embodiments, agents that reduce the level and / or activity of BRG1 and / or BRM include TALENs or mRNAs encoding TALENs that target (e.g., cut) nucleic acid sequences (e.g., DNA sequences) of BRG1. In embodiments, an agent that decreases the level and / or activity of BRG1 and / or BRM comprises a TALEN that targets (eg, cleaves) a nucleic acid sequence (eg, a DNA sequence) of a BRM or an mRNA encoding a TALEN.

[0231] For example, gRNA can be used in a CRISPR system to engineer changes in genes (e.g., BRG1 and / or BRM). In other embodiments, ZFN and / or TALEN can be used to engineer changes in genes (e.g., BRG1 and / or BRM). Exemplary changes include insertions, deletions (e.g., knockouts), translocations, inversions, single point mutations or other mutations. The changes can be introduced into genes in cells, for example, in vitro, ex vivo or in vivo. In some embodiments, the changes reduce (e.g., knock down or knock out) the level and / or activity of BRG1 and / or BRM, for example, the changes are negative function regulators. In another embodiment, the changes correct defects in BRG1 and / or BRM (e.g., mutations that cause defects). In another embodiment, the changes correct defects in BRG1 (e.g., mutations that cause defects). In another embodiment, the changes correct defects in BRM (e.g., mutations that cause defects).

[0232] In certain embodiments, the CRISPR system is used to edit (e.g., add or delete base pairs) target genes, such as BRG1 and / or BRM. In other embodiments, the CRISPR system is used to introduce premature stop codons, for example, so as to reduce the expression of the target gene. In other embodiments, the CRISPR system is used to turn off the target gene in a reversible manner (e.g., similar to RNA interference). In embodiments, the CRISPR system is used to guide Cas to the promoter of the target gene (e.g., BRG1 and / or BRM), thereby spatially blocking RNA polymerase. In embodiments, the CRISPR system is used to guide Cas to the promoter of the target gene (e.g., BRG1), thereby spatially blocking RNA polymerase. In embodiments, the CRISPR system is used to guide Cas to the promoter of the target gene (e.g., BRM), thereby spatially blocking RNA polymerase.

[0233] In some embodiments, a CRISPR system can be generated to edit BRG1 and / or BRM using techniques described in, for example, U.S. Publication No. 20140068797; Cong et al., Science 339(6121):819-823 (2013); Tsai, Nature Biotechnol., 32(6):569-576 (2014) and U.S. Pat. Nos. 8,871,445; 8,865,406; 8,795,965; 8,771,945 and 8,697,359.

[0234] In some embodiments, CRISPR interference (CRISPRi) technology can be used for transcriptional repression of specific genes (e.g., genes encoding BRG1 and / or BRM). In CRISPRi, the designed Cas9 protein (e.g., dCas9 without nuclease, or dCas9 fusion protein, such as dCas9-KRAB or dCas9-SID4X fusion) can be paired with sequence-specific guide RNA (sgRNA). The Cas9-gRNA complex can block RNA polymerase, thereby interfering with transcriptional extension. The complex can also block transcriptional initiation by interfering with transcription factor binding. The CRISPRi method is specific, has minimal off-target effects, and is multiple, for example, more than one gene (e.g., using multiple gRNAs) can be repressed simultaneously. In addition, the CRISPRi method allows reversible gene repression.

[0235] In some embodiments, CRISPR-mediated gene activation (CRISPRa) can be used for transcriptional activation, such as one or more genes described herein, such as genes that inhibit BRG1 and / or BRM. In CRISPRa technology, dCas9 fusion proteins recruit transcriptional activators. For example, dCas9 can be used to recruit polypeptides (e.g., activation domains) such as VP64 or p65 activation domains (p65D) and used with sgRNA (e.g., single sgRNA or multiple sgRNAs) to activate one or more genes, e.g., endogenous genes. Multiple activators can be recruited by using multiple sgRNAs, which can improve activation efficiency. A variety of activation domains and single or multiple activation domains can be used. In addition to engineering dCas9 to recruit activators, sgRNAs can also be engineered to recruit activators. For example, RNA aptamers can be incorporated into sgRNAs to recruit proteins (e.g., activation domains), such as VP64. In some instances, the synergistic activation mediator (SAM) system can be used for transcriptional activation. In SAM, an MS2 aptamer is added to the sgRNA. MS2 recruits MS2 coat protein (MCP) fused to p65AD and heat shock factor 1 (HSF1). CRISPRi and CRISPRa technologies are described in more detail, for example, in Dominguez et al., Nat. Rev. Mol. Cell Biol. 17(1): 5-15 (2016), which is incorporated herein by reference.

[0236] Small molecule compounds

[0237] In some embodiments of the invention, the agent that reduces the level and / or activity of BRG1 and / or BRM in a cell is a small molecule compound. In some embodiments, the small molecule compound is a structure of Formula I-III.

[0238] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula I or a pharmaceutically acceptable salt thereof:

[0239]

[0240] Where m is 0, 1, 2, 3 or 4;

[0241] X 1 is N or CH; and

[0242] Each R 1 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino.

[0243] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula II or a pharmaceutically acceptable salt thereof:

[0244]

[0245] Where R 2 is phenyl substituted by hydroxy and optionally substituted by one or more groups independently selected from the group consisting of halo, cyano, trifluoromethyl, trifluoromethoxy, C 1-3 Alkyl and C 1-3 Alkoxy;

[0246] R 3 Selected from the group consisting of: -R a 、-OR a 、-N(R a )2, -S(O)2R a and -C(O)-N(R a )2;

[0247] Each R a are independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl, wherein each C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6The alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of: R b , oxo, halogen, -NO2, -N(R b )2、-CN、-C(O)-N(R b )2、-S(O)-N(R b )2、-S(O)2-N(R b )2.-OR b 、-SR b 、-OC(O)-R b 、-C(O)-R b 、-C(O)-OR b 、-S(O)-R b 、-S(O)2-R b 、-N(R b )-C(O)-R b 、-N(R b )-S(O)-R b 、-N(R b )-C(O)-N(R b )2 and -N(R b )-S(O)2-R b ;

[0248] Each R b are independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, 3-15 membered carbocyclic group and 3-15 membered heterocyclic group, wherein each C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted by one or more independently selected from R c or two R b Together with the nitrogen to which they are attached, they form a heterocyclic group, which is optionally substituted by one or more independently selected from oxo, halo and C 1-3 alkyl, which is optionally substituted with one or more groups independently selected from the group consisting of oxo and halo;

[0249] Each Rc is independently selected from the group consisting of: oxo, halo, -NO2, -N(R d )2、-CN、-C(O)-N(R d )2、-S(O)-N(R d)2. -S(O) 2- N(R d )2. -SR d 、-OC(O)-R d 、-C(O)-R d 、-C(O)-OR d 、-S(O)-R d 、-S(O)2-R d 、-N(R d )-C(O)-R d 、-N(R d )-S(O)-R d 、-N(R d )-C(O)-N(R d )2、-N(R d )-S(O)2-R d , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl, any C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 The alkynyl, 3-15 membered carbocyclyl and 3-15 membered heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of: R d , oxo, halogen, -NO2, -N(R d )2、-CN、-C(O)-N(R d )2、-S(O)-N(R d )2、-S(O)2-N(R d )2.-OR d 、-SR d 、-OC(O)-R d 、-C(O)-R d 、-C(O)-R d 、-S(O)-R d 、-S(O)2-R d 、-N(R d )-C(O)-R d 、-N(R d )-S(O)-R d 、-N(R d )-C(O)-N(R d )2 and -N(R d )-S(O)2-R d ;

[0250] Each R d are independently selected from the group consisting of: hydrogen, C 1-6Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, carbocyclic and carbocyclic (C 1-3 alkyl)-;

[0251] R 4 It is H, C 1-6 Alkyl or -C(=O)-C 1-6 Alkyl; and

[0252] R 5 Is H or C 1-6 alkyl.

[0253] The compound of formula II can be synthesized by methods known in the art, such as those described in U.S. Patent Publication No. 2018 / 0086720, the synthesis methods of which are incorporated by reference.

[0254] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of Formula III or a pharmaceutically acceptable salt thereof:

[0255]

[0256] Where R 6 is halo, such as fluoro or chloro;

[0257] R 7 is hydrogen, optionally substituted amino or optionally substituted C 1-6 Alkyl; and

[0258] R 8 is an optionally substituted C 6-10 Aryl or optionally substituted C 2-9 Heteroaryl.

[0259] In some embodiments, the small molecule BRG1 and / or BRM inhibitor is a compound having the structure of any one of Compounds 1-16, or a pharmaceutically acceptable salt thereof:

[0260]

[0261]

[0262] In some embodiments, the small molecule compound or a pharmaceutically acceptable salt thereof is a degradation agent. In some embodiments, the degradation agent has a structure of Formula IV:

[0263] ALB

[0264] Formula IV

[0265] Wherein A is a BRG1 and / or BRM binding moiety; L is a linker; and B is a degradation moiety, or a pharmaceutically acceptable salt thereof. In some embodiments, the degradation moiety is a ubiquitin ligase moiety. In some embodiments, the ubiquitin ligase binding moiety comprises a Cereblon ligand, an IAP (inhibitor of apoptosis) ligand, a mouse double minute 2 homolog (MDM2), a hydrophobic tag, or a von Hippel-Lindau ligand or a derivative or analog thereof.

[0266] In some embodiments, A is a BRG1 binding moiety. In some embodiments, A is a BRM binding moiety. In some embodiments, A comprises the structure of any one of Formulas I-III or any one of Compounds 1-16.

[0267] In some embodiments, the hydrophobic tag comprises diphenylmethane, adamantane, or tri-Boc arginine, i.e., the hydrophobic tag comprises the following structure:

[0268]

[0269] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula A:

[0270]

[0271] Where X 1 is CH2, O, S or NR 1 , where R 1 X is H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 2 is C=O, CH2 or R 3 and R 4 is independently H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 heteroalkyl; m is 0, 1, 2, 3, or 4; and each R 2 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino,

[0272] or a pharmaceutically acceptable salt thereof.

[0273] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0274]

[0275] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0276] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula B:

[0277]

[0278] Each R 4 , R 4’ and R 7 R is independently H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 5 is an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R 6 is H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; n is 0, 1, 2, 3 or 4; each R 8 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino; and each R 9 and R 10 are independently H, halogen, optionally substituted C1-C6 alkyl or optionally substituted C6-C 10 Aryl, wherein R4' or R5 includes a bond to the linker, or a pharmaceutically acceptable salt thereof.

[0279] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0280]

[0281] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0282] In some embodiments, the ubiquitin ligase binding moiety comprises the structure of Formula C:

[0283]

[0284] Each R 11 , R 13 and R 15 R is independently H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl; 12 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; R 14 is an optionally substituted C1-C6 alkyl, an optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; p is 0, 1, 2, 3 or 4; each R 16 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino; q is 0, 1, 2, 3 or 4; and each R 17 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0285] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0286]

[0287] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0288] In some embodiments, the ubiquitin ligase binding moiety comprises a structure of Formula D:

[0289]

[0290] Each R 18 and R 19 are independently H, optionally substituted C1-C6 alkyl, optionally substituted C3-C 10 Carbocyclic group, optionally substituted C6-C 10 Aryl, optionally substituted C1-C6 alkyl, C3-C 10 Carbocyclic or optionally substituted C1-C6 alkyl C6-C 10 Aryl; r1 is 0, 1, 2, 3 or 4; each R 20 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxyl, thiol or optionally substituted amino; r2 is 0, 1, 2, 3 or 4; and each R 21 are independently halogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C 10 carbocyclyl, optionally substituted C2-C9 heterocyclyl, optionally substituted C6-C 10 aryl, optionally substituted C2-C9 heteroaryl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 heteroalkenyl, hydroxy, thiol or optionally substituted amino, or a pharmaceutically acceptable salt thereof.

[0291] In some embodiments, the ubiquitin ligase binding moiety comprises the following structure:

[0292]

[0293] or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0294] In some embodiments, the linker has the structure of Formula V:

[0295] A 1 -(B 1 ) f-(C 1 ) g -(B 2 ) h -(D)-(B 3 ) i -(C 2 ) j -(B 4 ) k -A 2

[0296] Formula V

[0297] Among them A 1 is the bond between the connector and A; A 2 is the bond between B and the joint; B 1 , B 2 , B 3 and B 4 are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2, and NRN; RN is hydrogen, optionally substituted C 1-4 Alkyl, optionally substituted C 2-4 Alkenyl, optionally substituted C 2-4 Alkynyl, optionally substituted C 2-6 Heterocyclic group, optionally substituted C 6-12 Aryl or optionally substituted C 1-7 Heteroalkyl; C 1 and C 2 are each independently selected from carbonyl, thiocarbonyl, sulfonyl or phosphoryl; f, g, h, I, j and k are each independently 0 or 1; and D is an optionally substituted C 1-10 Alkyl, optionally substituted C 2-10 Alkenyl, optionally substituted C 2-10 Alkynyl, optionally substituted C 2-6 heterocyclic group, optionally substituted and C 6-12 Aryl, optionally substituted C2-C 10 Polyethylene glycol or optionally substituted C 1-10 Heteroalkyl, or connected to A 1 -(B 1 ) f -(C 1 ) g -(B 2 ) h - to - (B 3 ) i -(C 2 ) j -(B 4 ) k -A 2 chemical bonds.

[0298] In some embodiments, D is optionally substituted C2-C 10 In some embodiments, C 1 and C 2 Each is independently carbonyl or sulfonyl. 1 , B 2 , B 3 and B 4 are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2 and NR N RN is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 , B 2 , B 3 and B 4 Each is independently selected from optionally substituted C1-C2 alkyl or optionally substituted C1-C3 heteroalkyl. In some embodiments, j is 0. In some embodiments, k is 0. In some embodiments, j and k are each independently 0. In some embodiments, f, g, h and i are each independently 1.

[0299] In some embodiments, the linker of Formula V has the structure of Formula Va:

[0300]

[0301] Among them A 1 is the bond between the linker and A, and A 2 It is the bond between B and the linker.

[0302] In some embodiments, D is optionally substituted C 1-10 In some embodiments, C 1 and C 2 In some embodiments, B 1 , B 2 , B 3 and B 4 are each independently selected from optionally substituted C1-C2 alkyl, optionally substituted C1-C3 heteroalkyl, O, S, S(O)2 and NR N , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 , B 2 , B 3 and B 4 Each independently selected from optionally substituted C1-C2 alkyl, O, S, S(O)2 and NRN , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 1 and B 4 Each is independently an optionally substituted C1-C2 alkyl. 1 and B 4 Each is independently C1 alkyl. 2 and B 4 Each is independently NR N , where R N is hydrogen or optionally substituted C 1-4 In some embodiments, B 2 and B 4 In some embodiments, f, g, h, I, j, and k are each independently 1.

[0303] In some embodiments, the linker of Formula V has the structure of Formula Vb:

[0304]

[0305] Among them A 1 is the bond between the linker and A, and A 2 It is the bond between B and the linker.

[0306] Preferably, the compound used in the methods disclosed herein has the following structure:

[0307]

[0308] or a pharmaceutically acceptable salt thereof.

[0309] More preferably, the compound used in the methods disclosed herein has the following structure:

[0310]

[0311] or a pharmaceutically acceptable salt thereof.

[0312] Combination preparations and uses thereof

[0313] An agent that reduces the level and / or activity of BRG1 and / or BRM can be combined with one or more therapeutic agents. In particular, the therapeutic agent can be a therapeutic agent for the treatment or prophylactic treatment of any of the cancers described herein.

[0314] Combination therapy

[0315] Agents that reduce the level and / or activity of BRG1 and / or BRM can be used alone, or in combination with another therapeutic agent (e.g., other agents for treating cancer or symptoms associated therewith), or in combination with other types of treatments for treating cancer. In combination therapy, the dose of one or more therapeutic compounds can be reduced from the standard dose when administered alone. For example, the dose can be determined empirically based on drug combinations and permutations, or can be derived by isobologram analysis (e.g., Black et al., Neurology 65: S3-S6, 2005). In this case, the dose of the compound should provide a therapeutic effect when combined.

[0316] In some embodiments, the second therapeutic agent is a chemotherapeutic agent (e.g., a cytotoxic agent or other compound useful for treating cancer). These include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione-substituted ureas, methylhydrazine derivatives, adrenocortical suppressants, adrenocortical steroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and gonadotropin-releasing hormone analogs. Also included are 5-fluorouracil (5-FU), folinic acid (LV), irinotecan, oxaliplatin, capecitabine, paclitaxel, and docetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide Alkyl sulfonates, such as busulfan, improsulfan and piposulfan; aziridines, such as benzodopa, carboquinone, mitedopam and euridopam; ethyleneimines and methylmelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; polyacetyl groups (especially bratacin and bratacinone); camptothecins (including the synthetic analogue topotecan); bryostatin; spongostatin; CC-1065 (including its synthetic analogues adolesine, carzelesine and biszelesine); nostoc (especially nostoc 1 and nostoc 8); pyraclostrobin; duocarmycins (including synthetic analogues, KW-218 9 and CB1-TM1); eleutherobin; mandinaine; stolonol; spongistatin; nitrogen mustards, such as chlorambucil, naphthiazine, clofosamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nembixin, phenamicin, prednimustine, trofosamide and uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicins, especially calicheamicin gamma II and calicheamicin omega II (see, e.g., Agnew, Chem. Intl. Ed. Engl. 33: 183-186 (1994); danemycins, including danemycin A; bisphosphonates such as clodronate; esperamicins; and the neocarcinogens and related chromoprotein enediyne antibiotic chromophores), aclarubicin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin C, carrubicin, carminomycin, carmophorin, chromomycin, dactinomycin, daunomycin, detoxibacin, 6-diazo-5-oxo-L-norleucine, (doxorubicin, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrroline-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, mexilomycin, mitomycin (such as mitomycin C), mycophenolic acid, noramycin, olive mycin, peplomycin, porfibrinocin, puromycin, triferric doxorubicin, rhodorubicin, streptozocin, streptozocin, tuberculocin, ubenimex, zoloft, doxycycline; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as folinic acid, methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine glycosides, doxifluridine, enocitabine, floxuridine; androgens, such as caprotestosterone, drostanolone propionate, cyclothiocarbamide, melastane, testolactone; antiadrenergic drugs, such as aminoglutethimide, mitotane, trilostan; folic acid supplements, such as folinic acid; aceglucuronolide; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; betribucil; bisantrene; edatrexate; defofamin; colchicine amine; diazocone; eflornithine; elliptinium acetate; epothilone; etoglu; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansine alkaloids, such as maytansine and ansamitocin; mitoguanidine; mitoxantrone; mopidarol; nitraerine; pentostatin; methamine nitrogen mustard; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; Polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizoran; spirogermanamine; tenuisporic acid; triazoline quinone; 2,2′,2″-trichlorotriethylamine; trichothecenes (particularly T-2 toxin, verrucosporin A, baculosporin A, and serpentin); urethane; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulanol; pipobroman; gacytosine; cytarabine (“Ara-C”); cyclophosphamide; thiotepa; taxanes, such as Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), Albumin-engineered paclitaxel nanoparticle formulation without Cremophor (American Pharmaceutical Partners, Schaumberg, Ill.) and docetaxel (Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum coordination complexes, such as cisplatin, oxaliplatin, and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; Vinorelbine; Noanto; Teniposide; Edatrexate; Daunomycin; Aminopterin; Xeloda; Ibandronate; Irinotecan (e.g., CPT-11); Topoisomerase inhibitor RFS2000; Difluoromethylornithine (DMFO); Retinoids, such as retinoic acid; Capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Two or more chemotherapeutic agents may be used in a mixture for administration in combination with the first therapeutic agent described herein. Suitable dosing regimens for combination chemotherapy are known in the art and are described, for example, in Saltz et al. (1999) Proc ASCO 18: 233a and Douillard et al. (2000) Lancet 355: 1041-7.

[0317] In some embodiments, the second therapeutic agent is a therapeutic agent that is a biologic used for cancer treatment, such as a cytokine (e.g., an interferon or an interleukin (e.g., IL-2)). In some embodiments, the biologic is an anti-angiogenic agent, such as an anti-VEGF agent, for example, bevacizumab In some embodiments, the biologic is an immunoglobulin-based biologic, such as a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Such agents include Rituxan (rituximab); Zenapax (daclizumab); Simulect (basiliximab); Synagis (palivizumab); Remicade (infliximab); Herceptin (trastuzumab); Mylotarg (gemtuzumab ozogamicin); Campath (alemtuzumab); Zevalin (ibritumomab tiuxetan); Humira (adalimumab); Xolair (omalizumab); Bexxar (tositumomab-I-131); Raptiva (efalizumab); Erbitux (cetuximab); Avastin (bevacizumab); Tysabri (natalizumab); Actemra (tositumomab); The invention also includes the following: (i) trastuzumab (Azar); (ii) trastuzumab (Azar); (iii) trastuzumab (Azar); (iv) trastuzumab (Azar); (iv) trastuzumab (Azar); (vectibix (panitumumab); (vientis (ranibizumab); (vientis (eculizumab); (vientibix (panitumumab); (vientis (ranibizumab); (vientibix (panitumumab); (vientis (ranibizumab); (vientibix (eculizumab); (vientibix (certolizumab); (iv) trastuzumab (Azar ...

[0318] The second agent can be a non-drug therapeutic agent. For example, the second therapeutic agent is radiation therapy, cryotherapy, hyperthermia and / or surgical removal of tumor tissue.

[0319] The second agent can be a checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody, such as a monoclonal antibody). The antibody can be, for example, humanized or fully human. In some embodiments, the checkpoint inhibitor is a fusion protein, for example, an Fc-receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent that interacts with a checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is an agent that interacts with a ligand of a checkpoint protein, such as an antibody. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA4 antibody, such as ipilimumab / Yervoy or tremelimumab). In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or a small molecule inhibitor) of PD-1 (e.g., nivolumab / Pembrolizumab / Pidilizumab ( / CT-011). In some embodiments, the checkpoint inhibitor is an inhibitor of PDL1 (e.g., an inhibitory antibody or a small molecule inhibitor) (e.g., MPDL3280A / RG7446; MEDI4736; MSB0010718C; BMS 936559). In some embodiments, the checkpoint inhibitor is an inhibitor of PDL2 (e.g., an inhibitory antibody or Fc fusion or a small molecule inhibitor) (e.g., PDL2 / Ig fusion protein, such as AMP 224). In some embodiments, the checkpoint inhibitor is an inhibitor of B7-H3 (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7 family ligands, or a combination thereof (e.g., an inhibitory antibody or a small molecule inhibitor).

[0320] In some embodiments, the agent that reduces the level and / or activity of BRG1 and / or BRM is used in combination with another anticancer therapy (e.g., surgery, MEK inhibitors and / or PKC inhibitors, or a combination thereof). For example, in some embodiments, the method further includes performing surgery before, after, or simultaneously with the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM. In some embodiments, the method further includes administering a MEK inhibitor (e.g., selumetinib, bimetinib, or tametinib) and / or a PKC inhibitor (e.g., sotrastaurin or IDE196) before, after, or simultaneously with the administration of an agent that reduces the level and / or activity of BRG1 and / or BRM.

[0321] In any combination embodiment described herein, the first therapeutic agent and the second therapeutic agent are administered simultaneously or sequentially in any order. The first therapeutic agent can be administered immediately, at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 5 hours, at most 6 hours, at most 7 hours, at most 8 hours, at most to 9 hours, at most 10 hours, at most 11 hours, at most 12 hours, at most 13 hours, 14 hours, at most 16 hours, at most 17 hours, at most 18 hours, at most 19 hours, at most 20 hours, at most 21 hours, at most 22 hours, at most 23 hours, at most 24 hours, or at most 1-7, 1-14, 1-21, or 1-30 days before or after the second therapeutic agent.

[0322] Pharmaceutical composition

[0323] The compounds described herein can be formulated into pharmaceutical compositions for administration to human subjects in a biocompatible form suitable for in vivo administration. Pharmaceutical compositions generally include an active agent as described herein and a physiologically acceptable excipient (e.g., a pharmaceutically acceptable excipient). The formulation principles of the compounds disclosed herein may be, for example, those described in WO 2020 / 160180, the disclosures of which are incorporated herein by reference in their entirety.

[0324] The compounds of the present invention can be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration, and the pharmaceutical composition is formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal and topical administration modes. Parenteral administration can be performed by continuous infusion in a selected time period. Preferably, the compound is administered orally.

[0325] Suitable pharmaceutical carriers as well as pharmaceutical requisites for pharmaceutical formulations are described in the well-known reference texts in the art, Remington: The Science and Practice of Pharmacy, 21st edition, Gennaro ed., Lippencott Williams & Wilkins (2005) and USP / NF (United States Pharmacopeia and National Formulary).

[0326] Unit dosage form

[0327] The compounds described herein may be formulated in unit dosage forms (eg, capsules) for oral administration.

[0328] The unit dosage form can contain suitable pharmaceutical carriers and excipients as described in the pharmaceutical composition section. Excipients can include, for example, anti-adherents, antioxidants, adhesives, coatings, compression aids, disintegrants, dyes (pigments), softeners, emulsifiers, fillers (diluents), film formers or coatings, flavorings, spices, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, and water for hydration. The unit dosage form can contain one or more of fillers, disintegrants, wetting agents, glidants, lubricants, and capsule shells.

[0329] In some embodiments, the filler may be 70% to 90% (w / w) of the unit dosage form. The filler may be microcrystalline cellulose, mannitol, or a combination thereof.

[0330] In some examples, the disintegrant may be 4% to 6% (w / w) of the unit dosage form.The disintegrant may be croscarmellose sodium.

[0331] In some examples, the wetting agent may be 0.5% to 1.5% (w / w) of the unit dosage form. The wetting agent may be sodium lauryl sulfate.

[0332] In some examples, the glidant may be 1.5% to 2.5% (w / w) of the unit dosage form. The glidant may be colloidal silicon dioxide.

[0333] In some examples, the lubricant may be 0.4% to 0.6% (w / w) of the unit dosage form. The lubricant may be magnesium stearate.

[0334] In some examples, the capsule shell is made of a polymer shell. The polymer shell can be made of hypromellose and titanium dioxide.

[0335] The compounds described herein can be formulated into unit dosage forms (e.g., capsules) for oral administration as described in Table 1. The API in Table 1 is a compound of the following structure:

[0336]

[0337] Table 1

[0338]

[0339] The composition of the Swedish Orange HPMC capsule shell is described in Table 2.

[0340] Table 2

[0341]

[0342] The composition of the blue-green HPMC capsule shell is described in Table 3.

[0343] Table 3

[0344]

[0345] Example

[0346] A multi-dose escalation clinical trial of the compound shown below is conducted. It is primarily intended to evaluate the safety and tolerability of the compound shown below when administered orally to subjects with advanced hematological malignancies, particularly relapsed or refractory acute myeloid leukemia (R / R AML) or myelodysplastic syndrome (MDS).

[0347]

[0348] In this ongoing study, one of 21 subjects with acute myeloid leukemia treated with the compounds shown above has been reported to have developed clinically significant signs and symptoms related to blood cell differentiation. This occurred approximately 7-14 days after treatment with the compounds shown above.

[0349] Other Implementations

[0350] Although the invention has been described in conjunction with specific embodiments thereof, it will be appreciated that it is capable of further modifications and that this application is intended to cover any changes, uses or adaptations that generally follow the principles of the invention and that are within the known or customary practice in the art to which the invention pertains and that may be applied to the key features indicated hereinabove and that follow the scope of the claims. Other embodiments are in the claims.

Claims

1. A method of treating a subject having clinically significant signs and symptoms associated with blood cell differentiation and being treated with an agent that reduces the level and / or activity of BRG1 and / or BRM, the method comprising administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

2. A method of treating a subject having differentiation syndrome and being treated with an agent that reduces the level and / or activity of BRG1 and / or BRM, the method comprising administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

3. A method of treating a subject suspected of having differentiation syndrome and being treated with an agent that reduces the level and / or activity of BRG1 and / or BRM, the method comprising administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

4. The method of any one of claims 1 to 3, wherein the subject is treated with an effective amount of the agent that reduces the level and / or activity of BRG1 and / or BRM of the leukemia.

5. The method of claim 4, wherein the leukemia is acute myeloid leukemia.

6. The method of any one of claims 1 to 3, wherein the subject is treated with an effective amount of the agent that reduces the level and / or activity of BRG1 and / or BRM of myelodysplastic syndrome.

7. The method of any one of claims 1 to 5, wherein the method further comprises administering the agent that reduces the level and / or activity of BRG1 and / or BRM.

8. A method of treating a subject having leukemia or myelodysplastic syndrome, the method comprising administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM, and if the subject has clinically significant signs and symptoms associated with blood cell differentiation, administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

9. A method of treating a subject having leukemia or myelodysplastic syndrome, the method comprising administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM, and if the subject has differentiation syndrome, administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

10. A method of treating a subject having leukemia or myelodysplastic syndrome, the method comprising administering an effective amount of an agent that reduces the level and / or activity of BRG1 and / or BRM, and if the subject is suspected of having differentiation syndrome, administering to the subject an effective amount of a corticosteroid, hydroxyurea or furosemide, or performing leukapheresis on the subject.

11. The method of any one of claims 8 to 10, wherein the subject has leukemia.

12. The method of claim 11, wherein the leukemia is acute myeloid leukemia.

13. The method of any one of claims 8 to 10, wherein the subject has myelodysplastic syndrome.

14. The method of any one of claims 1 to 13, wherein the subject exhibits one or more of the following symptoms: unexplained fever, rash, hypoxia, respiratory impairment, interstitial lung infiltrates, pleural and / or pericardial effusions, weight gain, renal failure, dyspnea, clinical deterioration, fluid in or around the lungs, fluid around the heart, leg swelling, increased bilirubin, and increased liver enzymes.

15. The method of any one of claims 1 to 14, wherein the blood sample from the subject comprises an elevated absolute neutrophil count (ANC) and / or an elevated platelet count.

16. The method of any one of claims 1 to 15, wherein an effective amount of a corticosteroid is administered to the subject.

17. The method of claim 16, wherein the corticosteroid is administered systemically.

18. The method of claim 16, wherein the corticosteroid is administered orally or by injection.

19. The method of any one of claims 16 to 18, wherein a high-dose regimen of a corticosteroid is administered to the subject.

20. The method of any one of claims 16 to 19, wherein the corticosteroid is administered to the subject for at least 3 days.

21. The method of any one of claims 16 to 20, wherein the corticosteroid is dexamethasone, inxamethasone, hydrocortisone, cortisone, prednisone, prednisolone, methylprednisolone, triamcinolone, a pharmaceutically acceptable salt thereof, or a combination thereof.

22. The method of any one of claims 16 to 21, wherein administration of the agent that reduces the level and / or activity of BRG1 and / or BRM is discontinued if the clinically significant signs and symptoms associated with blood cell differentiation or symptoms of the differentiation syndrome persist for at least 48 hours after initiation of administration of a corticosteroid.

23. The method of any one of claims 16 to 21, wherein administration of the agent that reduces the level and / or activity of BRG1 and / or BRM is discontinued if the clinically significant signs and symptoms associated with blood cell differentiation or the symptoms of the differentiation syndrome persist for at least 3 days after initiation of administration of a corticosteroid.

24. The method of any one of claims 1 to 23, wherein the subject has symptoms of non-infectious leukocytosis.

25. The method of any one of claims 1 to 24, wherein the method comprises administering to the subject an effective amount of hydroxyurea.

26. The method of claim 25, wherein an effective amount of hydroxyurea is administered to the subject until the non-infectious leukocytosis improves or resolves.

27. The method of any one of claims 1 to 26, wherein the method comprises performing leukapheresis on the subject.

28. The method of any one of claims 1 to 27, wherein the subject experiences hypervolemia.

29. The method of claim 28, wherein an effective amount of furosemide is administered to the subject.

30. The method of any one of claims 1 to 29, wherein the agent that reduces the level and / or activity of BRG1 and / or BRM is a compound of the structure: or a pharmaceutically acceptable salt thereof.

31. The method of any one of claims 1 to 29, wherein the agent that reduces the level and / or activity of BRG1 and / or BRM is a compound of the structure: or a pharmaceutically acceptable salt thereof.

32. The method of any one of claims 1 to 31, wherein the agent that reduces the level and / or activity of BRG1 and / or BRM is administered orally.

33. The method of any one of claims 1 to 32, wherein the agent that reduces the level and / or activity of BRG1 and / or BRM is administered in a unit dosage form selected from the group consisting of a capsule or a tablet.

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